Cellular Structure Visualize cell junctions and cellular adhesion Antibodies to study specific protein localization and cellular processes Cellular adhesion is essential for providing physical support and positional information in multicellular organisms. Such functions are mediated by adhesion receptor recognition of extracellular matrix (or cellular receptors on other cells) and through recep- tor control of cytoskeletal architecture and signaling cascades (Figure 1). Adhesion receptor signal transduction also integrates with signals from other receptor systems to regulate cell move- ment, differentiation, proliferation, and survival. Deregulation of adhesion contributes to disease progression, either by disrupt- ing the normal docking and movement of cells, which alters regulatory signaling, or by promoting inappropriate temporal Figure 1—Detection of Pyk2 [pY402] in NMµMG (mouse mammary) cells stimu- and spatial adhesion. lated with 20 ng/ml TGFβ for 36 hours. Pyk2 [pY402] was identified and visual- Cell junctions influence both the structure and behavior ized using BioSource™ Pyk2 [pY402]–specific antibody directly conjugated of adjacent cells by regulating how these cells interact with to Alexa Fluor® 488 dye (Invitrogen Cat. no. 44-618A1). Nuclei are stained each other and with the extracellular matrix. Understanding cell with DAPI (blue). The inset image shows cells not treated with TGFβ (control), junctions and their role in triggering cellular adhesion signaling which lack Pyk2 phosphorylation at site 402. pathways is fundamental to understanding disease processes, and for developing treatments that target tumor plasticity and angiogenesis. Unfortunately, it is difficult to discretely visualize cellular junctions and their associated signaling cascades, due in part to challenges in finding primary antibodies that recognize their targets with high specificity and affinity. Now you can get highly specific antibodies that allow you to visualize the pro- teins and protein modifications that are critical to your research (Figure 2). Zymed® antibodies for the study of cell junctions, and BioSource™ antibodies for cell adhesion research (both avail- Figure 2—Indirect immunofluorescence staining of adherens junctions between able from Invitrogen), produce highly specific identification of highly confluent HeLa cells using Zymed® mouse anti–α-catenin (Invitrogen target proteins. You’ll detect only the proteins you want, with Cat. no. 13-9700). α-Catenin (green) was visualized using goat anti–mouse IgG Alexa Fluor® 488 (Invitrogen Cat. no. A11029). Slides were mounted in low background. Invitrogen’s cell junction and cell adhesion ProLong® Gold Antifade Reagent (Invitrogen Cat. no. P36931), which con- antibodies are widely used and widely cited throughout the lit- tains DAPI for nuclear counterstaining (blue). F-actin (red) was visualized erature, demonstrating their reliability. using Alexa Fluor® 635 phalloidin (Invitrogen Cat. no. A34054). Get the whole picture. To learn more about Invitrogen’s antibodies for the study of cell junctions and cellular adhesion, visit www.invitrogen.com/antibodies. www.invitrogen.com ©2007 Invitrogen Corporation. All rights reserved. These products may be covered by one or more Limited Use Label Licenses (see Invitrogen catalog or www.invitrogen.com). By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. For research use only. Not intended for any animal or human therapeutic or diagnostic use, unless otherwise stated. A-070271-r1 US 0607 Amazing! Over 4,000 high quality antibodies from a single source Sigma’s AMAZING collection of high quality antibodies: Q Includes monoclonals, polyclonals, secondaries, conjugates and custom services Q Validated using a variety of methods – WB, IHC, IF, IP, ELISA Q Easily explored through Sigma’s award-winning online tools Q Supported by Sigma’s world-renowned technical service to assist you with your Don’t miss this Amazing specifi c antibody requirements opportunity: Order your copy of the all new 2007-2008 Sigma is your source. Antibodies Catalog, visit Go to sigma.com/antibody and be Amazed! sigma.com/antibody_cat Our Innovation, Your Research — Shaping the Future of Life Science sigma.com INNOVATIVE DISCOVERY TOOLS FOR SIGNAL TRANSDUCTION RESEARCH Over 300 Antibodies Validated for Flow Cytometry...from Cell Signaling Technology Phospho-Stat5 (Tyr694) (C11C5) Phospho-Histone H3 (Ser10) Antibody Rabbit mAb #9359 AID (EK2 5G9) Rat mAb #4959 (Alexa Fluor® 488 Conjugate) #9708 Flow cytometric analysis of K562 cells, untreated Flow cytometric analysis of Raji cells using #4959 Flow cytometric analysis of THP-1 cells treated with (green) or STI571-treated (blue), using #9359. (blue) compared to a nonspecific negative control paclitaxel using #9708 versus propidium iodide (DNA antibody (red). content). The box indicates phospho-histone H3 positive cells. :: Highest quality antibodies from CST provide the brightest signal with the lowest background. :: Extensive validation by our in-house clinical applications group — optimization is not left up to you, the user. :: Technical support provided by our clinical applications group — this translates into a thorough, fast and accurate response. :: Rapidly expanding line of Alexa Fluor® conjugated primary antibodies available for use in flow cytometry. Alexa Fluor ® is a registered trademark of Molecular Probes, Inc. For a complete product listing please visit... www.cellsignal.com Orders (toll-free) 1-877-616-CELL (2355) [email protected] Fax ordering 1-978-867-2488 Technical support (toll-free) 1-877-678-TECH (8324) [email protected] Inquiries [email protected] In flow cytometry, every dot tells a story . The ImageStream® system HL60 cells were labeled with AF488 anti- HLA to reveal the cell membrane and with DRAQ5TM to stain the nucleus. Labeled cells were analyzed on the ImageStream sys- tem. The DNA content histogram (above) cleanly separates the major cell cycle subpopulations. G2/M cells were plotted (right) to display the high nuclear texture fraction, from which cells in the progressive VWDJHVRIPLWRVLVZHUHLGHQWLÀHGDQGKLJK- lighted on the plot. Composite cell images (far right) show HLA and nuclear staining. F r A q t S c m UNTRANSLOCATED TRANSL l W s c t T ImageStream. Think outside the dot. Amnis®, ImageStream® and IDEAS® are registered trademarks of Amnis Corporation. DRAQ5 is a trademark of Biostatus, Ltd. CELL BIOLOGY CELL SIGNALING DRUG DISCOVERY IMMUNODETECTION CLEAR SIGNALS? LAB WATER PROTEIN BIOMARKERS Understand what’s happening along the pathway. STEM CELL RESEARCH We help accelerate the progress you’re making in Life Science, by providing the most comprehensive support and advanced selection of tools to help drive your cell signaling research forward. We partner with leaders in the fi eld to make the latest discoveries available for you. And, with the Upstate portfolio of products and expertise, including the largest offering of modifi cation state-specifi c antibodies, we have the solutions for all of your research needs. ADVANCING LIFE SCIENCE TOGETHER THE EXPERTISE OF ® Visit www.millipore.com for more information on this and other ways UPSTATE Millipore supports Life Science research. IS NOW A PART OF MILLIPORE 7V^LYM\S[VVSZ[V LS\JPKH[L[OLILHZ[ CD25 CD4 GITR CD46 CD28 CD45RO CD137 CD278 (ICOS) *OLJRV\[V\Y M\SSSPULVM (U[PIVKPLZHUK 9LJVTIPUHU[ 7YV[LPUZMVY/\THU 0TT\UVSVN`9LZLHYJO H[www.ancell.com Ancell Corporation Human CD4+ 5 Day PHA Blasts ;OPYK:[YLL[5VY[OQ76)V_Q)H`WVY[45<:( ;VSS-YLL! ;LS! -H_! HUJLSS'HUJLSSJVTQ^^^HUJLSSJVT Europe:(UJLSS,\YVWLJV(SL_PZ*VYWVYH[PVU 0UK\Z[YPLZ[YHZZLQ7VZ[MHJOQ*/3H\ZLU:^P[aLYSHUK ;LS! -H_! HSL_PZJO'HSL_PZJVYWJVTQ^^^HSL_PZJVYWJVT anti-GITR/R-PE Japan:*VZTV)PV*V3[KQ;VR`V ;LS! Q-H_! ^^^NSVILVYQWJVZTVIPV anti-ICOS/FITC International Distributors:Austria:,<)06 Australia:0TT\UV+PHNUVZ[PJZ Bangladesh:-<;<9,)<:05,::=0:065Belgium:7»Z)=)(Canada: 0+3():PUJVY -H_ 4,+0*69705* VY-H_Czech Republic: .,5,;0*(:96Denmark:4,+056=(:*0,5;0-0*(: Europe: (5*,33,<967,JV(3,?0:*69769(;065 Finland:5<77<3055(53()69(;69067(3=,3<6@ France: *6=(3()*6.,9:(Germany: (??69(+,<;:*/3(5+.TI/ Greece:)(3*(54,+ :))06;,*/5636.@:<7730,9:(,Hong Kong: )677(9+/2*63;+ Hungary:)064(92,93;+ India:/@:,305+0(7][3[KIndonesia:0;:05+65,:0( Ireland:(37/(;,*/5636.0,:3;+Israel: (346.+0(.56:;0* 4,+0*(3,8<074,5;3;+ Italy:=05*0)06*/,4Japan:*6:46)06*63;+ 0>(0*OLTPJHSZ*V3[K Malaysia:05;,9:*0,5*,:+5)/+ New Zealand: :(77/09,)06:*0,5*,7;@3;+ Norway:(/+0(.56:;0*:(:Pakistan: ;/,>693+>0+,:*0,5;0-0*Poland: )0640)6 Portugal: )(7;0:;(4(98<,:3+(PR China: +HRL^L)PV[LJO*V Romania: 4,+0:;:( Russia: */044,+05* Singapore: 0;::*0,5*, 4,+0*(37;,3;+ South Korea: *695,3)06;,*/*697 9LZLHYJO5,;0UJ Spain: .9<76;(7,9:( Sweden: 2,3()Switzerland: (3,?0:*69769(;065 Taiwan:.LU/V[3HIVYH[VYPLZ0UJ Thailand:0;:;/(03(5+*63;+The Netherlands:7»ZTurkey: )PV.LU ;629(4,+0*(33;+ UK and Republic of Ireland: (??69(<23;+ Try the new Human FoxP3 Clone 259D/C7 Introducing the new human FoxP3 (clone 259D/C7) and simple buffer system from BD Biosciences. 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