Immunofluorescent analysis of Heat Shock Protein 60 (Hsp60) in U251 Cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a Heat Shock Protein 60 (Hsp60) monoclonal antibody (Product # MA3-013) at a dilution of 1:200 overnight at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35503). Heat Shock Protein 60 (Hsp60) staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.
|Tested species reactivity||Human, Mouse, Non-human primate, Rat|
|Published species reactivity||Not Applicable|
|Host / Isotype||Mouse / IgM|
|Immunogen||Human placental HSP60.|
|Storage buffer||1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ELISA (ELISA)||1-10 µg/ml|
|Immunohistochemistry (IHC)||1-10 µg/ml|
|Immunoprecipitation (IP)||2 µg|
|Western Blot (WB)||1-10 µg/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|
MA3-013 detects HSP60 from human, monkey, mouse, and rat samples. This antibody displays no other protein reactivity.
MA3-013 has been successfully used in Western blot, immunoprecipitation, immunohistochemistry, immunofluorescence, and ELISA procedures. By Western blot, this antibody detects a single 58 kDa protein representing recombinant human HSP60.
The MA3-013 antigen is human placental HSP60. Epitope mapping studies using human HSP60 deletion mutants suggest that this antibody binds between amino acids 211-288.
Reconstitute with PBS.
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. HSP60, an ~60 kDa protein, is the human homologue of HSP65 (a major antigenic mycobacterial protein) and E. coli GroEL. HSP60, also referred to as chaperonin-60, P1 and mitonin, has been classified as a member of a family of proteins termed chaperonins which act to recognize and stabilize polypeptide intermediates during folding, assembly and disassembly. HSP60 is a constitutively expressed mitochondrial protein that shares many characteristics with HSP70 including high abundance, induction by environmental stress, ATPase activity and a critical role in protein assembly and folding. However, in contrast with HSP70, HSP60 functions as a 14 subunit oligomer which is thought to interact with only 1 or 2 target polypeptides at one time.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Primary structure of a human mitochondrial protein homologous to the bacterial and plant chaperonins and to the 65-kilodalton mycobacterial antigen.
MA3-013 was used in western blot to clone and characterize human hsp60
|Jindal S,Dudani AK,Singh B,Harley CB,Gupta RS||Molecular and cellular biology (9:2279)||1989|