Product Specification Sheet HeLa cells (normal/non-heat shocked) total lysate for western control for hsps

 Cat. # HSP703-C Human HeLa cells (normal/non-heat shocked) total protein lysate for western control for hsps SIZE: 100 ul

 Cat. # HSP704-C Human HeLa cells (Heat shocked) total protein lysate for western control for hsps SIZE: 100 ul

Heat shock , as a class, are among the most highly dissolved in 1X SDS-PAGE Sample buffer (reduced) for direct expressed cellular proteins across all species. Heat shock proteins loading on gels for Western. Total protein concentration is 10 ug/10 protect cells when stressed by elevated temperatures. They account ul. For Western blot +ve control are supplied in SDS-PAGE sample for 1–2% of total protein in unstressed cells and up to 4–6% of total buffer (reduced). Load 10 ul/lane of lysates for good visibility with protein in stressed cells.. The 70 kilodalton heat shock proteins antibody Cat # HSP702-M or # HSP703-A or other antibodies to () are a family of ubiquitously expressed heat shock proteins. . Store at –20oC in suitable size aliquots. SDS may Members of the Hsp70 family are strongly upregulated by heat crystallize in cold conditions. It should redissolve by warming before stress and toxic chemicals, particularly heavy metals such as taking it from the stock. It should be heated once prior to loading on arsenic, cadmium, copper, mercury, etc. HSP 70 is overexpressed gels. If the product has been stored for several weeks, then it may in malignant melanoma[10] and underexpressed in renal cell cancer. be preferable to add 5 ul of fresh 2x sample buffer per 10 ul of the lysate solution prior to heating and loading on gels. This preparation The Hsp70 proteins have three major functional domains: is not biologically active. It is not suitable for ELISA or other N-terminal ATPase domain – binds ATP and hydrolyzes it to ADP. applications where native protein is required. Do not freeze, thaw, or The exchange of ATP drives conformational changes in the other heat repeatedly. two domains. Substrate binding domain – contains a groove with Fig: Total protein profile of HeLa an affinity for neutral, hydrophobic amino acid residues. C-terminal (normal, lane 2) and heat shocked domain – rich in alpha helical structure acts as a 'lid' for the (lane 3). The two lanes on the right substrate binding domain. were probed with anti-hsp70 anitbody #HSP702-M. The arrow Prokaryotes express three Hsp70 proteins: DnaK, HscA (Hsc66), indicates the hsp70 positions and HscC (Hsc62). Eukaryotic organisms express several slightly (induced) after heat shock. different Hsp70 proteins. All share the common domain structure, but each has a unique pattern of expression or subcellular localization. These are, among others:

Hsc70 (Hsp73/HSPA8) is a constitutively expressed protein. It typically makes up one to three percent of total cellular protein. Hsp70 (encoded by three very closely related paralogs: HSPA1A, HSPA1B, and HSPA1L) is a stress-induced protein. High levels can Form & Storage of be produced by cells in response to hyperthermia, oxidative stress, Storage and changes in pH. O Binding immunoglobulin protein (BiP or Grp78) is a protein localized Short-term: unopened, undiluted liquid vials at -20 C and powder at 4OC or -20OC. to the endoplasmic reticulum. It is involved in there, O and can be upregulated in response to stress or starvation. Long-term: at –20 C or below in suitable aliquots after mtHsp70 or Grp75 is the mitochondrial Hsp70. reconstitution. Do not freeze and thaw and store working, diluted solutions.

Source of Antigen, Antibodies O Stability: 6-12 months at –20 C or below. The HeLa cell line was originally derived from a human Shipping: 4OC for solutions and room temp for powder. donor's epithelioid carcinoma tissue (cervix). Heat shocking reproducibly alters protein expression in cells, resulting in Recommended Usage various morphological changes and the expression of heat shock proteins crucial for both maintaining cellular Western Blot: load 5-10 ul per lane of HeLa protein lysates.. homeostasis during normal cell growth and for survival during and after various cellular stresses. In mammalian General References: Flaherty KM (1990) Nature 346, 623-628; cells, the induction of the heat shock response requires the Tavaria M (1996) Cell Stress Chaperones 1, 23-28; Morano KA activation and nuclear translocation of transcription factors (2007) Ann. NY Acad. Sci. 1113, 1-14; Yoshimune K (2002) BBRC which control the expression of cytoprotective heat shock 293, 1389-1395. proteins, and play pivotal roles in the development of inflammation and immunity. Normal and heat-shocked cell lysates offers a convenient way to study heat-shock related *This product is for In vitro research use only. protein changes.

#HSP703-C and #HSP704-C Related Items Anti-, Hsp90 (Acetylated at K69, K100, K294, K558) Normal HeLa cells (HSP703-C) are grown at 37oC and heat- Recombinant hsp90 alpha and beta isoforms shocked cells (#HSP704-C) are exposed to 42oC for 30 min before cell harvesting in log phase. Cells are washed, and the cell pellet is HSP703-C 140109A

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