Immunofluorescent analysis of Heat Shock Protein 90 (HSP90, green) in HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed without (left panel) or with (right panel) an HSP90 monoclonal antibody (Product # MA3-010) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight 554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan or a ToxInsight Instrument at 20X magnification.
|Tested species reactivity||Bovine, Human, Mouse, Rat|
|Published species reactivity||Rabbit, Non-human primate, Bovine, Mouse, Human|
|Host / Isotype||Mouse / IgG2b|
|Immunogen||Purified mouse HSP90.|
|Storage buffer||ascites diluted in PBS|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20|
|Western Blot (WB)||1:500|
* 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.
MA3-010 detects heat shock protein 90 kDa (HSP90) from human, mouse, rat, and bovine tissues. This antibody does not detect chicken HSP90.
MA3-010 has been successfully used in Western blot, immunofluorescence, and immunohistochemistry (paraffin) procedures. By Western blot, this antibody detects a 90 kDa protein representing HSP90 from MCF7 cell extract. MA3-011 is more sensitive in Western blot procedures and should be used when maximum sensitivity is desired. MA3-010 does not immunoprecipitate HSP90 in any form (free or complexed).
The MA3-010 antigen is purified mouse HSP90.
Heat shock proteins (HSP) are proteins that are expressed in response to various biological stress, including heat. HSP90 is a 90 kDa protein that is induced under stress conditions, but is also one of the most abundant cellular proteins found under non-stress conditions. HSP90 has been found to be associated with a number of other intracellular proteins, including steroid receptors, actin, tubulin, and some kinases. It has been suggested that HSP90 is necessary to maintain structural integrity of at least some steroid receptors to allow for ligand binding during receptor activation.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
The distinct agonistic properties of the phenylpyrazolosteroid cortivazol reveal interdomain communication within the glucocorticoid receptor.
MA3-010 was used in western blot to study the mechanism for differential actions of various synthetic steroids on glucocorticoid receptor.
|Yoshikawa N,Yamamoto K,Shimizu N,Yamada S,Morimoto C,Tanaka H||Molecular endocrinology (Baltimore, Md.) (19:1110)||2005|
Effect of stages of lactation on the concentration of a 90-kilodalton heat shock protein in bovine mammary tissue.
MA3-010 was used in western blot to investigate the differences of HSP90 expression in cow mammary tissue at various lactation stages
|Watanabe A,Miyamoto T,Katoh N,Takahashi Y||Journal of dairy science (80:2372)||1997|
Agonist-free transformation of the glucocorticoid receptor in human B-lymphoma cells.
MA3-010 was used in western blot to study the role of cell-cell interactions in the ligand-independent nuclear translocation of the glucocorticoid receptor
|van den Berg JD,Smets LA,van Rooij H||The Journal of steroid biochemistry and molecular biology (57:239)||1996|
High levels of non-activated receptors in glucocorticoid-sensitive S49wt mouse lymphoma cells incubated with dexamethasone.
MA3-010 was used in western blot to investigate the glucocorticoid receptor status of murine S49 lymphoma cells exposed to dexamethasone
|van den Berg JD,Smets LA,Hutchison KA,van Rooij H,van den Elshout MM||The Journal of steroid biochemistry and molecular biology (51:33)||1994|
ATP-dependent protein refolding activity in reticulocyte lysate. Evidence for the participation of different chaperone components.
MA3-010 was used in western blot to investigate the role of chaperone components for energy-dependent protein refolding activity
|Nimmesgern E,Hartl FU||FEBS letters (331:25)||1993|
Oolemmal proteomics--identification of highly abundant heat shock proteins and molecular chaperones in the mature mouse egg and their localization on the plasma membrane.
MA3-010 was used in immunohistochemistry to identify highly abundant heat shock proteins and chaperone proteins in mature mouse egg proteome.
|Calvert ME,Digilio LC,Herr JC,Coonrod SA||Reproductive biology and endocrinology : RB and E (1:null)||2003|