Western blot analysis of Heat Shock Protein 70 (Hsp70) was performed by loading 30ug of THP-1 cell lysate per well onto an SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 1 hour at room temperature. The membrane was probed with an Hsp70 monoclonal antibody (Product # MA1-10889) at a dilution of 1:1000 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated goat anti-mouse IgG secondary antibody at a dilution of 1:40,000 for 1 hour at room temperature. Chemiluminescent detection was performed using ECL substrate. Data courtesy of the Innovators Program.
|Tested species reactivity||Many|
|Published species reactivity||Not Applicable|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Human Hsp70 purified from HeLa cells.|
|Storage buffer||PBS, pH 7.4, with 50% glycerol|
|Contains||0.1% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ELISA (ELISA)||1-10 ug/ml|
|Flow Cytometry (Flow)||10 ug/ml|
|Immunohistochemistry (IHC)||5-10 ug/ml|
|Immunoprecipitation (IP)||1-10 ug/ml|
|Western Blot (WB)||1 ug/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Miscellaneous PubMed (MISC)||See 1 publications below|
MA1-10889 detects Heat Shock Protein 70 in human, mouse, rat, rabbit, monkey, guinea pig, hamster, chicken, bovine, canine, ovine, porcine, fish, Drosophila and C. elegans samples and does not cross-react with Hsc70.
MA1-10889 has been successfully used in Western blot, immunohistochemistry, immunoprecipitation, ELISA and flow cytometry procedures. By Western blot, MA1-10889 is specific for the ~70 kDa Heat Shock Protein 70 protein.
MA1-10889 immunogen is human Hsp70 purified from HeLa cells, corresponding to amino acids 436-503.
Heat shock proteins (HSP) are expressed in response to various biological stresses, including high temperatures. There are several major families of HSPs including HSP70, HSP90 and HSP100. The HSP70 family is a set of highly conserved proteins that are induced by a variety of biological stresses, including heat stress, in every organism in which the proteins have been examined. The human HSP70 family members include: HSP70, a protein which is strongly inducible in all organisms but which is also constitutively expressed in primate cells; HSP72, a 72 kDa protein that is induced exclusively under stress conditions; HSC70, or cognate protein, is a 72 kDa, constitutively expressed, protein which is involved in the uncoating of clathrin coated vesicles; GRP78, or BiP, is a glucose regulated 78 kDa protein localized in the endoplasmic reticulum; and p75, or HSP75, a 75 kDa protein that is found within the mitochondria.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Cell cycle-dependent association of HSP70 with specific cellular proteins.
MA1-10889 was used in immunocytochemistry, immunoprecipitation and western blot to investigate the association of HSP70 and specific cellular proteins during the cell cycle
|Milarski KL,Welch WJ,Morimoto RI||The Journal of cell biology (108:413)||1989|