LEPTOMYCIN A+B Inhibitors of Nuclear-Cytoplasmic Transport Highlighthighlight Tomorrow’S Reagents Manufactured Today™ International Version

PRODUCT FLYER LEPTOMYCIN A+B Inhibitors of Nuclear-Cytoplasmic Transport highlighthighlight Tomorrow’s Reagents Manufactured Today™ International Version

Available Nuclear Transport at Best Prices! In eukaryotic cells, the enclosure of ge- 65nm long aqueous channel. Transport oc- netic information by the nucleus allows the curs by a variety of different pathways, de- spatial and temporal separation of DNA rep- fined by individual receptors and accessory Leptomycin A+B lication and transcription in the nucleus from soluble factors. Soluble factors selectively cytoplasmic protein synthesis. This compart- target substrates/cargo for import and ex- mentalization permits a high level of regula- port as well as deliver them to their appropri- Leptomycin A [LMA] tion of these processes, but at the same time ate intracellular destination. Macromolecules ALX-380-101-C050 50 µg necessitates a system of selective macromo- (cargo) transported through these channels ALX-380-101-C100 100 µg lecular transport between the nucleus and include proteins and RNA, as individual enti- An antifungal antibiotic that acts as an inhibitor cytoplasm [1, 2]. ties or as part of larger complexes such as the of nuclear export by interacting with CRM1/ex- The term nuclear-cytoplasmic transport ribosomal subunits. While small molecules portin-1. Inhibits nucleo-cytoplasmic transloca- such as ions and proteins of up to 60kDa tion of molecules such as the HIV-1 Rev protein refers to the movement of a large variety of and Rev-dependent export of mRNA. macromolecules both into and out of the can diffuse through the NPC, small proteins nucleus. actively cross the NPC in a carrier-mediated SOURCE: Isolated from Streptomyces sp. fashion [3, 4]. FORMULATION: In 100% ethanol. A single type of channel, the nuclear Proteins to be imported into the nucle- MW: 526.7. FORMULA: C H O pore complex (NPC), mediates all move- 32 46 6 ment across the nuclear envelope. The NPC us contain sequences termed nuclear locali- PURITY: ≥95% (HPLC). CAS: 87081-36-5 is a massive structure, completely spanning zation sequences (NLS), and proteins to be For Bulk Quantities please inquire! the two membranes that separate the nucle- exported from the nucleus contain nuclear export sequences (NES). These signals are re- Manufactured by Novartis AG for us from the cytoplasm and housing a central Dolder AG Switzerland

Leptomycin B [LMB] ALX-380-100-C100 100 µg An antifungal antibiotic that acts as an inhibitor of nuclear export by interacting with CRM1/ exportin-1. Inhibits nucleo-cytoplasmic translocation of molecules such as the HIV-1 Rev protein and Rev-dependent export of mRNA. Other proteins that are influenced by leptomycin B are actin, c-Abl, cyclin B1, MDM2/p53, Iκb, MPF and PKA. SOURCE: Isolated from Streptomyces sp. FORMULATION: In 100% ethanol.

MW: 540.7. FORMULA: C33H48O6 PURITY: ≥95% (HPLC). CAS: 87081-35-4 For Bulk Quantities please inquire!

Leptomycin Set I ALX-850-313-KI01 1 Set The Set contains 1 x 100µg each of Leptomycin A (Prod. No. ALX-380-101) and Leptomycin B (Prod. No. ALX-380-100). FIGURE 1: Classical nuclear-cytoplasmic transport 2 IMMUNOLOGYLEPTOMYCIN A+B

cognized by soluble factors that work with the RanGTPase cycle to coordinate import or export Leptomycin B: A Well Established Standard from nucleus [5, 6]. Leptomycin B (LMB) is an antibiotic with of many proteins including HIV-1 Rev (the hu- Protein import process: The NLS is a protein anti-fungal and anti-tumor activity that was man immunodeficiency virus type 1 regula- motif containing a cluster of basic residues that first discovered and purified from the fermen- tory protein), MAPK/ERK, and NF-κB/IκB. Ad- is recognized by soluble adapter proteins known tation broth and mycelia of Streptomyces. LMB ditionally, it also stabilizes the expression of as importins. The importin proteins, also termed (C33H48O6) is an unsaturated, branched chain p53 by protecting p53 from Mdm2-mediated karyopherins (Table 1), is one member of a con- fatty acid with a terminal lactone ring (Figure degradation and inducing p53 transcriptional served family of transport receptors. Each mem- 3). activity. Leptomycin B also inhibits the export ber of this conserved family recognizes a distinct Later, this drug was shown to inhibit the G1 and translation of many RNAs, including COX- NLS. Through a cascade of associations with oth- 2 and c-Fos mRNAs, by inhibiting export of ri- er components within the nuclear translocation and G2 phases of the cell cycle, and other cellular functions. These pleotropic effects led to bonucleoproteins. Other proteins that have process, the importins ultimately interact with been shown to be influenced by leptomycin the phenylalanine-glycine (FG) repeat regions the study of the mechanisms by which leptomycin B influences intracellular processes. One B, include actin, c-Abl, cyclin B1, MPF, PKA and of the nucleoporins (a protein subunit of the nu- many others [20-29]. clear pore complex) and with the cofactor Ran- of these studies involved a screen of leptomy- GTP (the GTP-bound form of the RanGTPase) to cin B resistance genes in fission yeast. In this Export of mRNA and certain proteins from discharge the cargo in the nucleus. Alternatively, screen it was found that leptomycin B targets the nucleus is a key step in protein produc- an importin-independent pathway also exists. In CRM1 (exportin-1), a fission yeast nuclear pro- tion, proliferation, and apoptosis. Therefore, this case, the cargos contain several HEAT-like tein, which was thought to be involved in the this antibiotic has become an important tool repeats and can associate directly with the nu- control of gene expression [16-19]. for studying nuclear localization and traffick- cleo-porins [5-10]. Leptomycin B forms a covalent complex ing in eukaryotic cells. Mechanism for nuclear export: Studies of with the sulfhydryl group of a conserved the HIV Rev and the cellular PKI (protein kinase cysteine residue in CRM1, thereby inhibiting A inhibitor) proteins led to the discovery of the CRM1 interaction with the nuclear export sig- leucine-rich nuclear export signal (NES) and the nal (NES) of targeted export proteins. The abil- very well characterized CRM1/exportin-1 (chro- ity of leptomycin B to inhibit nuclear export has made it a useful tool in the study of the mosome region maintenance 1) pathway of nu- FIGURE 3 clear export. CRM1 is a karyopherin specific for subcellular localization of many regulatory pro- nuclear export (exportin 1). The CRM1 protein di- teins. Leptomycin B blocks the nuclear export rectly binds proteins that contain a leucine-rich NES. In addition, the export mechanism appears to involve CRM1 binding to both Ranγ-GTP and nucleoporins (Figure 1) [11-14]. Leptomycin A: The Unexplored Alternative! Nuclear-cytoplasmic transport is an impor- Leptomycin A (LMA) (Figure 2) is an antifun- to the lack of supply and high prices, LMA is tant aspect of normal cell function. Defects in gal antibiotic that inhibits nucleo-cytoplasmic still unexplored. this process have been detected in many differ- translocation of Rev and MAPK/ERK, by inhib- ent types of cancer cells. These defects can occur iting their export. LMA belongs to a defined in the signal transduction pathways that regulate panel of nuclear export inhibitors, including the transfer of factors such as p53 and β-catenin leptomycin and ratjadones. Although most in and out of the nucleus, or in the general nucle- of research done on leptomycin was done on us import and export machinery itself [15]. LMB, the specifications for LMA indicate simil- iar blocking potential of the nuclear export FIGURE 2 system by leptomycin A [16-18, 30, 31]. Due

Literature References

1 Nucleocytoplasmic transport: taking an inventory: H. Fried and U. Kutay; Cell. Mol. Life Sci. 60, 1659 (2003) (Review) 2 Nuclear Func- tions for Plasma Membrane-Associated Proteins?: A. Benmerah, et al.; Traffic 4, 503 (2005) (Review) 3 The nuclear pore complex: a protein machine bridging the nucleus and cytoplasm: K.J. Ryan and S.R. Wente; Curr. Opin. Cell Biol. 12, 361 (2000) (Review) 4 Gatekeepers of the nucleus: S.R. Wente; Science 288, 1374 (2000) (Review) 5 Leucine-rich nuclear-export signals: born to be weak: U. Kutay and S. Guttinger; Trends Cell Biol. 15, 121 (2005) (Review) 6 Mechanisms of Recptor-Mediated Nuclear Import and Nuclear Export: L.F. Pemberton and B.M.

Paschal; Traffic 6, 187 (2005) (Review) 7 Importin alpha: a multipurpose nuclear-transport receptor: D.S. Goldfarb, et al.; Trends Cell Biol. 14, 505 (2004) (Review) 8 Importin beta: conducting a much larger cellular symphony: A. Harel and D.J. Forbes; Mol. Cell 16, 319 (2004) (Review) 9 Karyopherins: from nuclear-transport mediators to nuclear-function regulators: N. Mosammaparast and L.F. Pemberton; Trends Cell Biol. 14, 547 (2004) (Review) 10 Nucleocytoplasmic shuttling of signal transducers: L. Xu and J. Massague; Nat. Rev. Mol. Cell Biol. 5, 209 (2004) (Review) 11 CRM1 is an export receptor for leucine-rich nuclear export signals: M. Fornerod, et al.; Cell 90, 1051 (1997) 12 CRM1 is responsible for intracellular transport mediated by the nuclear export signal: M. Fukuda, et al.; Nature 390, 308 (1997) 13 Evidence for a role of CRM1 in signal-mediated nuclear protein export: B. Ossareh-Nazari, et al.; Science 278, 141 (1997) 14 Exportin 1 (Crm1p) is an essential nuclear export factor: K. Stade, et al.; Cell 90, 1041 (1997) 15 Nuclear transport and cancer: from mechanism to intervention: T.R. Kau, et al.; Nat. Rev. Cancer 4, 106 (2004) (Review) 16 Leptomycins A and B, new antifungal antibiotics. I. Taxonomy of the producing strain and their fermentation, purification and characterization: T. Hamamoto, et al.; J. Antibiot. (Tokyo) 36, 639 (1983) 17 Leptomycins A and B, new antifungal antibiotics. II. Structure elucidation: T. Hamamoto, et al.; J. Antibiot. (Tokyo) 36, 646 (1983) 18 Leptomycins A and B, new antifungal antibiotics. III. Mode of action of leptomycin B on Schizosaccharomyces pombe: T. Hamamoto, et al.; J. Antibiot. (Tokyo) 38, 1573 (1985) 19 Effects of leptomycin B on the cell cycle of fibroblasts and fission yeast cells: M. Yoshida, et al.; Exp. Cell Res. 187, 150 (1990) 20 TABLE 1: Human Karyopherins Interaction of MAP kinase with MAP kinase kinase: its possible role in the control of nucleocytoplasmic transport of MAP kinase: M. Fukuda, et al.; Embo J. 16, 1901 (1997) 21 Leptomycin B is an inhibitor of nuclear export: inhibition of nucleo-cytoplasmic translocation of the human immunodeficiency virus type 1 (HIV-1) Rev protein and Rev-dependent mRNA: B. Wolff, et al.; Chem. Biol. 4, 139 (1997) 22 Nuclear export is required for degradation of endogenous p53 by MDM2 and human papillomavirus E6: D.A. Freedman and A.J. Levine; Mol. Cell Biol. 18, 7288 Selected Latest Review Articles (1998) 23 Loss of IkappaB alpha-mediated control over nuclear import and DNA binding enables oncogenic activation of c-Rel: S. Sachdev and M. Hannink; Mol. Cell Biol. 18, 5445 (1998) 24 Nuclear export of MAP kinase (ERK) involves a MAP kinase kinase (MEK)-dependent active Regulation of nuclear transport: central role in development transport mechanism: M. Adachi, et al.; J. Cell Biol. 148, 849 (2000) 25 Protein ligands to HuR modulate its interaction with target mRNAs in and transformation?: I.K. Poon and D.A. Jans; Traffic 6, 173 vivo: C.M. Brennan, et al.; J. Cell Biol. 151, 1 (2000) 26 Alteration of poly(ADP-ribose) glycohydrolase nucleocytoplasmic shuttling character- (2005) istics upon cleavage by apoptotic proteases: M.E. Bonicalzi, et al.; Biol. Cell 95, 635 (2003) 27 Leptomycin B, an inhibitor of the nuclear export Mechanisms of receptor-mediated nuclear import and nuclear receptor CRM1, inhibits COX-2 expression: B.C. Jang, et al.; J. Biol. Chem. 278, 2773 (2003) 28 Nuclear export inhibitor leptomycin B induces export: L.F. Pemberton and B.M. Paschal; Traffic 6, 187 (2005) the appearance of novel forms of human Mdm2 protein: S. Menendez, et al.; Br. J. Cancer 88, 636 (2003) 29 Hypoxia induces p53 through Translocation through the nuclear pore complex: selectivity a pathway distinct from most DNA-damaging and stress-inducing agents: A. Renton, et al.; Carcinogenesis 24, 1177 (2003) 30 Biosynthetic and speed by reduction-of-dimensionality: R. Peters; Traffic 6, studies of leptomycins: T. Hamamoto, et al.; J. Antibiot. (Tokyo) 38, 533 (1985) 31 The unique C-terminal tail of the mitogen-activated pro- 421 (2005) tein kinase ERK5 regulates its activation and nuclear shuttling: M. Buschbeck and A. Ullrich; J. Biol. Chem. 280, 2659 (2005)

For updated prices and additional information visit www.alexis-biochemicals.com, contact your Local Distributor, or call +41 61 926 89 89 LEPTOMYCIN A+B 3

Inhibitors of Nuclear- Inhibitors Cytoplasmic Transport Ratjadone A (synthetic) BAPTA/AM The nuclear-cytoplasmic transport of pro- ALX-270-346-C002 2 µg ALX-450-014-M010 10 mg teins is a necessity for normal cell function. In Synthetic. Represents a new class of natural com- ALX-450-014-M050 50 mg the last few years the understanding of the pounds, which inhibit proliferation in eukaryotes by Cell permeable acetoxymethyl ester derivative of mechanism of transport into and out of the blocking nuclear export. As potent as leptomycin BAPTA (Prod. No. ALX-450-013). Useful for control- B and specific for G1/S checkpoint. Cytotoxic sec- 2+ nucleus and the identification of the main ling intracellular Ca concentration. Induces inacti- ondary metabolite (IC50=50pg/ml in mouse cell line vation of protein kinase C (PKC). Also inhibits thapsi- responsible factors (importins, exportins, L929) that arrests tumor cells in the G1 phase at re- gargin-induced apoptosis in rat thymocytes. nucleoporins and Ran regulators) has pro- markably low concentrations (50pg/ml in HeLa cell LIT: Physiological [Ca2+]i level and pump-leak turnover in intact gressed (see related products). The next chal- line KB3.1). Inhibits the binding between the nucle- red cells measured using an incorporated Ca chelator: V.L. Lew, et lenge is to understand the different mecha- ar export signal (NES) of proteins and the chromo- al.; Nature 298, 478 (1982) BAPTA induces a decrease of intracel- some maintenance region protein (CRM1). Antican- lular free calcium and a translocation and inactivation of protein nisms by which nuclear transport can be cer compound. Belongs to the family of orphan li- kinase C in macrophages: P. Dieter, et al.; Biol. Chem. Hoppe-Sey- regulated. gands which include polyketides like leptomycin B, ler 374, 171 (1993) The development of small molecule inhib- callystatin A and other related compounds. SN50 itors and the identification of natural product LIT: The chemistry and biology of ratjadone: M. Kalesse, et al.; Chem- BioChem. 9, 709 (2001) The chemistry and biology of the lepto- H-Ala-Ala-Val-Ala-Leu-Leu-Pro-Ala-Val-Leu-Leu-Ala-Leu-Leu- inhibitors of transport receptors, or specific mycin family: M. Kalesse & M. Christmann; Synthesis 8, 981 (2002) Ala-Pro-Val-Gln-Arg-Lys-Arg-Gln-Lys-Leu-Met-Pro-OH factors like nucleoporins are useful in under- Ratjadones inhibit nuclear export by blocking CRM1/exportin 1: M. ALX-167-024-C500 500 µg Koster, et al.; Exp. Cell. Res. 286, 321 (2003) standing transport pathways. Inhibits translocation of the NF-κB active complex Up to date, a selected panel of inhibito- into the nucleus. For control peptide, see SN50M (Prod. No. ALX-167-025). ry compounds was described and designed. LIT: Inhibition of nuclear translocation of transcription factor NF-ka- Important natural products in this field are ppa B by a synthetic peptide containing a cell membrane-permeable leptomycin A and B (see page 1 + 2), ratja- motif and nuclear localization sequence: Y.Z. Lin, et al.; J. Biol. Chem. 270, 14255 (1995) Nuclear factor-kappaB activates dual inhibition done A and C [1, 2] next to, callystatin A [3, 4], sites in the regulation of tumor necrosis factor-alpha-induced neu- thapsigargin [5], PKF050-638 [6], valtrate [7], trophil apoptosis: M. Niwa, et al.; Eur. J. Pharmacol 407, 211 (2000) wheat germ agglutinin (WGA) [8], and agari- Activation of nuclear factor kappaB and induction of apoptosis by Ratjadone C (native) tumor necrosis factor-alpha in the mouse uterine epithelial WEG-1 cus bisporus lectin (ABL) [9]. In addition, small ALX-270-369-C005 5 µg cell line: S. Pampfer, et al.; Biol. Reprod. 63, 879 (2000) chemical or proteonic molecules were identified to have an inhibitory effect on nuclear Isolated from Sorangium cellulosum. Cytotoxic sec- SN50M Control Peptide ondary metabolite that inhibits cell growth of mam- transport like p10 protein [10], GTPγS [11], H-Ala-Ala-Val-Ala-Leu-Leu-Pro-Ala-Val-Leu-Leu-Ala-Pro-Val- malian cell lines in the picomolar range (IC50=0.2ng/ Gln-Arg-Asn-Gly-Gln-Lys-Leu-Met-Pro-OH BAPTA [11], truncated importin-β [12], SN50 ml with L929 mouse cells) including multidrug resist- ALX-167-025-C500 500 µg peptide and cell-permeant cyclized SN50 [13, ant HeLa cells (IC50=0.1ng/ml with KB-V1). Like lepto- 14] or N-ethylmaleide (NEM) [15, 16]. Beneath mycin B, ratjadone C binds covalently to the nuclear Inactive control peptide for SN50 (Prod. No. ALX- a panel of new compounds [17, 18] were syn- export protein CRM1. It inhibits cargo protein bind- 167-024). ing to the leucine-rich nuclear export sequence and LIT: Inhibition of nuclear translocation of transcription factor NF-ka- thesized, which were designed after eluci- thereby blocks nuclear export. ppa B by a synthetic peptide containing a cell membrane-perme- dation of the structural organisation of the LIT: Antibiotics from gliding bacteria, LXIII. Ratjadone: a new anti- able motif and nuclear localization sequence: Y.Z. Lin, et al.; J. Biol. transport system itself. fungal metabolite from Sorangium cellulosum: D. Schummer, et al.; Chem. 270, 14255 (1995) Liebigs Ann. 685 (1995) Ratjadon: a new antifungal compound from Sorangium cellulosum (myxobacteria) production, physico-chemical GTPγ S and biological properties: K. Gerth, et al.; J. Antibiot. (Tokyo) 48, 973 LIT: 1 The chemistry and biology of ratjadone: M. Kalesse, et al.; JBS-NU-412S 20 units Chembiochem. 2, 709 (2001) 2 Ratjadones inhibit nuclear ex- (1995) Ratjadones inhibit nuclear export by blocking CRM1/expor- port by blocking CRM1/exportin 1: M. Koster, et al.; Exp. Cell Res. tin 1: M. Koster, et al.; Exp. Cell. Res. 286, 321 (2003) JBS-NU-412L 100 units 286, 321 (2003) 3 The Chemistry and Biology of the Leptomycin LIT: The interferon-induced 67-kDa guanylate-binding protein Family: M. Kalesse; Synthesis 8, 981 (2002) 4 The total synthesis (hGBP1) is a GTPase that converts GTP to GMP: M. Schwemmle and of (-)-callystatin A: M. Kalesse, et al.; Chemistry 9, 1129 (2003) 5 P. Staeheli; J. Biol. Chem. 269, 11299 (1994) GTP-induced membrane Depletion of calcium from the lumen of endoplasmic reticulum binding and ion channel activity of annexin VI: is annexin VI a GTP reversibly inhibits passive diffusion and signal-mediated trans- biosensor?: A. Kirilenko, et al.; Biophys. J. 82, 2737 (2002) port into the nucleus: U.F. Greber and L. Gerace; J. Cell Biol. 128, 5 (1995) 6 A synthetic HIV-1 Rev inhibitor interfering with the Mant-GTPγS CRM1-mediated nuclear export: D. Daelemans, et al.; PNAS 99, 14440 (2002) 7 New Rev-transport inhibitor with anti-HIV activ- JBS-NU-209S 10 units ity from Valerianae Radix: N. Murakami, et al.; Bioorg. Med. Chem. JBS-NU-209L 50 units Lett. 12, 2807 (2002) 8 Inhibition of in vitro nuclear transport Thapsigargin LIT: Interdomain interactions regulate GDP release from heterot- by a lectin that binds to nuclear pores: D.R. Finlay, et al.; J. Cell Biol. ALX-350-004-M001 1 mg rimeric G proteins: A.E. Remmers and R.R. Neubig; Biochemistry 38, 104, 189 (1987) 9 Edible mushroom (Agaricus bisporus) lectin, ALX-350-004-M005 5 mg 13795 (1999) 2’(3’)-O-(N-methylanthraniloyl)-substituted GTP ana- which reversibly inhibits epithelial cell proliferation, blocks nucle- logs: a novel class of potent competitive adenylyl cyclase inhibitors: ar localization sequence-dependent nuclear protein import: L.G. ALX-350-004-M010 10 mg A. Gille and R. Seifert; J. Biol. Chem. 278, 12672 (2003) Yu, et al.; J. Biol. Chem. 274, 4890 (1999) 10 Role of the nuclear ALX-350-004-M025 25 mg transport factor p10 in nuclear import: U. Nehrbass and G. Blo- bel; Science 272, 120 (1996) 11 Assembly of the nuclear pore: Cell permeable tumor promoter by specific inhibi- biochemically distinct steps revealed with NEM, GTP gamma S, tion of the endoplasmatic reticulum Ca2+-ATPase. and BAPTA: C. Macaulay and D.J. Forbes; J. Cell Biol. 132, 5 (1996) Does not increase inositol phosphates. Shows no Now Available! 12 Dominant-negative mutants of importin-beta block mul- effect on protein kinase C (PKC). Increases Ca2+-de- tiple pathways of import and export through the nuclear pore + Ratjadones and Leptomycins are structurally simi- complex: U. Kutay, et al.; Embo J. 16, 1153 (1997) 13 Nuclear pendent Na influx in human platelets in a dose-de- import of proinflammatory transcription factors is required for pendent manner. Induces apoptosis. lar natural products, belonging to the “Leptomycin massive liver apoptosis induced by bacterial lipopolysaccharide: LIT: Thapsigargin, a Ca(2+)-ATPase inhibitor, depletes the intracellu- family” of orphan Ligands. Both compounds inhib- D. Liu, et al.; J. Biol. Chem. 279, 48434 (2004) 14 Suppression of lar Ca2+ pool and induces apoptosis in human hepatoma cells: A. its proliferation in eukaryotes by blocking nuclear Staphylococcal Enterotoxin B-induced Toxicity by a Nuclear Im- Tsukamoto & Y. Kaneko; Cell Biol. Int. 17, 969 (1993) export, exploring anti-cancer activities! port Inhibitor: D. Liu, et al.; J. Biol. Chem. 279, 19239 (2004) 15 An N-ethylmaleimide-sensitive cytosolic factor necessary for nuclear protein import: requirement in signal-mediated binding to BAPTA . tetrapotassium salt Leptomycin Set II the nuclear pore: D.D. Newmeyer and D.J. Forbes; J. Cell Biol. 110, ALX-450-013-G001 1 g ALX-850-316-KI01 1 Set 547 (1990) 16 A cytosolic activity distinct from crm1 mediates Highly selective Ca2+ chelating agent. The Set contains 1 x 100µg each of Leptomycin A nuclear export of protein kinase inhibitor in permeabilized cells: J.M. Holaska and B.M. Paschal; PNAS 95, 14739 (1998) 17 A LIT: New calcium indicators and buffers with high selectivity against (Prod. No. ALX-380-101) and Leptomycin B (Prod. chemical genetic screen identifies inhibitors of regulated nuclear magnesium and protons: design, synthesis, and properties of proto- No. ALX-380-100) and 1 x 5µg Ratjadone C (native) export of a Forkhead transcription factor in PTEN-deficient tumor type structures: R.Y. Tsien; Biochemistry 19, 2396 (1980) Transmitter (Prod. No. ALX-270-369). cells: T.R. Kau, et al.; Cancer Cell 4, 463 (2003) 18 Nuclear export release at frog end-plate loaded with a Ca2+-chelator, BAPTA: hyper- inhibitors and kinase inhibitors identified using a MAPK-activated tonicity and erythrosin B augment the release independently of in- For updated prices please visit protein kinase 2 redistribution screen: D.L. Almholt, et al.; Assay ternal Ca2+: N. Tanabe & H. Kijima; Neurosci. Lett. 92, 52 (1988) Drug Dev. Technol. 2, 7 (2004)

Purified (PF) = Purified (Preservative free); FC = Flow Cytometry; ICC = Immunocytochemistry; IP = Immunoprecipitation; IHC = Immunohistochemistry (FS = Frozen Sections, PS = Paraffin Sections); WB = Western blot; BP = Blocking Peptide 4 LEPTOMYCIN A+B

Nuclear Import/Export

PAb to Chromosome Segregation 1-like PAb to Importin (α2 Subunit) (1-50) Ran (G19V/Q69L mutant) (human) Protein (1-50) (human) (BL1975) (BL1959) (recombinant) BET-A300-472A Purified 0.1 mg BET-A300-483A Purified 0.1 mg JBS-PR-214 50 µg From rabbit. IMMUNOGEN: Synthetic peptide corre- From rabbit. IMMUNOGEN: Synthetic peptide corre- Recombinant Ran (G19V/Q69L) produced in E. coli. sponding to aa 1-50 of human CSE1L (chromosome sponding to aa 1-50 of human importin (α2 subunit). The Ran mutant G19V/Q69L is defective in GTP hy- segregation 1-like protein). SPECIFICITY: Recognizes SPECIFICITY: Recognizes human importin (α2 subu- drolysis. Substitution of glycine 19 by valine results human CSE1L. Cross-reactivity with other species nit). Cross-reactivity with other species has not been in a gain-of-function mutant that is not sensitive to has not been tested. APPLICATION: IP, WB. BP: BET- tested. APPLICATION: IP, WB. BP: BET-BP300-483. the exchange factor RCC1. BP300-472. PAb to Importin (α2 Subunit) (475-529) Ran (Q69L mutant) (human) PAb to Chromosome Segregation 1-like (BL1961) (recombinant) Protein (900-950) (BL1977) BET-A300-484A Purified 0.1 mg JBS-PR-213 50 µg BET-A300-473A Purified 0.1 mg From rabbit. IMMUNOGEN: Synthetic peptide corre- Recombinant Ran (Q69L) produced in E. coli. The Ran From rabbit. IMMUNOGEN: Synthetic peptide corre- sponding to aa 475-529 of C-terminal human impor- mutant Q69L is defective in GTP hydrolysis. Useful sponding to aa 900-950 of C-terminal human CSE1L tin (α2 subunit). SPECIFICITY: Recognizes human im- as a negative control for Ran binding and import (chromosome segregation 1-like protein). SPECIFICI portin (α2 subunit). Cross-reactivity with other spe- studies. TY: Recognizes human and mouse CSE1L. Cross-re- cies has not been tested. APPLICATION: IP, WB. BP: activity with other species has not been tested. AP BET-BP300-484. Ran (T24N mutant) (human) PLICATION: WB. BP: BET-BP300-473. PAb to Importin (β1 Subunit) (1-50) (recombinant) PAb to Exportin-1 (1025-1071) (BL1958) (BL1962) JBS-PR-212 50 µg BET-A300-469A Purified 0.1 mg BET-A300-481A Purified 0.1 mg Recombinant Ran (T24N) produced in E. coli. The Ran mutant T24N has no or little binding to GTP and is From rabbit. IMMUNOGEN: Synthetic peptide cor- From rabbit. IMMUNOGEN: Synthetic peptide corre- responding to aa 1025-1071 of C-terminal hu- β known to act as a negative inhibitor of nuclear im- sponding to aa 1-50 of importin ( 1 subunit). SPE port. man exportin-1 (chromosome region mainte- CIFICITY: Recognizes human and mouse importin (β1 nance 1; CRM1). SPECIFICITY: Recognizes human subunit). Cross-reactivity with other species has not Rna1p (S. pombe) (recombinant) and mouse exportin-1. Cross-reactivity with other been tested. APPLICATION: WB. BP: BET-BP300-481. species has not been tested. APPLICATION: IP, WB. JBS-PR-221 20 µg BP: BET-BP300-469. PAb to Importin (β1 Subunit) (825-876) Recombinant Rna1p (S. pombe) produced in E. coli. (BL1964) Rna1p is the GTPase-activating factor (GAP) of yeast PAb to Exportin-1 (375-425) (human) Gsp1p, an ortholog of mammalian Ran. Rna1p is lo- (BL1956) BET-A300-482A Purified 0.1 mg cated in the cytoplasm throughout the cell cycle and From rabbit. IMMUNOGEN: Synthetic peptide corre- plays a direct role in protein import into the nucleus BET-A300-490A Purified 0.1 mg sponding to aa 825-876 of C-terminal human impor- in S. cerevisiae and S. pombe. From rabbit. IMMUNOGEN: Synthetic peptide corre- tin (β1 subunit). SPECIFICITY: Recognizes human and sponding to aa 375-425 of human exportin-1 (chro- mouse importin (β1 subunit). Cross-reactivity with mosome region maintenance 1; CRM1). SPECIFICITY: other species has not been tested. APPLICATION: Recognizes human exportin-1. Cross-reactivity with IP, WB. BP: BET-BP300-482. other species has not been tested. APPLICATION: IP. BP: BET-BP300-490. MAb to Nuclear Transport Factor p97 Nuclear Pore Complex (3E9) GppNHp ALX-804-025-R100 100 µl JBS-NU-401S 100 units CLONE: 3E9. ISOTYPE: Mouse IgG2a. IMMUNOGEN: Pu- MAb to Nuclear Pore Glycoprotein JBS-NU-401L 500 units rified bovine NTF97 (nuclear transport factor p97). (O-Linked) (RL1) LIT: Structure of a Ran-binding domain complexed with Ran bound SPECIFICITY: Recognizes human, rat and dog NTF97. ALX-804-113-R100 100 µl to a GTP analogue: implications for nuclear transport: I.R. Vetter, et APPLICATION: ICC, IP, WB. FUNC: Inhibits import of NLS- al.; Nature 398, 39 (1999) containing proteins in permeabilized cells. MAb to N-Acetylglucosamine (O-linked) Importin α (human) (recombinant) (RL2) (His) Ran (E70A mutant) (human) (recombinant) ALX-804-111-R100 100 µl JBS-PR-231 50 µg JBS-PR-215 50 µg Recombinant human His-tagged importin α pro- MAb to Nucleoporin p62 (rat) (RL31) duced in E. coli. Recombinant Ran (E70A) produced in E. coli. The Ran ALX-804-275-R200 200 µl mutant E70A changed glutamic acid to alanine and LIT: Importins and exportins: how to get in and out of the nucleus: severly limited the guanine exchange reaction medi- K. Weis; TIBS 23, 185 (1998) Transport into and out of the nucleus: MAb to Nucleolar Protein 1 (yeast) I.G. Macara; Microbiol. Mol. Biol. Rev. 65, 570 (2001) ated by RCC1, the guanine nucleotide exchange fac- (28F2) tor (GEF) for Ran. Importin β (D. melanogaster) PTS-40-1236-R200 200 µl (recombinant) (His) MAb to Nucleolar Protein 2 (yeast) JBS-PR-232 50 µg (22G1) Recombinant His-tagged importin β (D. mela- PTS-40-1237-R200 200 µl nogaster) produced in E. coli. More Information? Please visit LIT: See JBS-PR-231 MAb to Nucleoporin NSP1 (yeast) (32D6) PTS-40-1239-R200 200 µl 1-Mar-06

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