Immunofluorescent analysis of Heat Shock Protein 84 using Anti-Heat Shock Protein 84 Polyclonal Antibody (Product# PA3-012) shows staining in U251 Cells. Heat Shock Protein 84 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Heat Shock Protein 84 (Product# PA3-012) at a dilution of 1:100 over night at 4 °C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product# 35552, Goat Anti-Rabbit). Images were taken at 60X magnification.
|Tested species reactivity||Human, Mouse, Non-human primate, Rat|
|Published species reactivity||Rat, Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues P(2) E E V H H G E E E V E(13) of mouse HSP84.|
|Purification||Antigen affinity chromatography|
|Storage buffer||1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1-20 µg/ml|
|Immunocytochemistry (ICC)||10-20 µg/ml|
|Immunofluorescence (IF)||10 - 20 µg/ml|
|Immunohistochemistry (Frozen) (IHC (F))||10 ug/ml|
|Immunoprecipitation (IP)||2 µg|
|Western Blot (WB)||1:1000 - 1:20000|
* 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.
PA3-012 detects heat shock protein 90 beta (HSP90) from human, monkey, mouse, and rat tissues and cells. This antibody does not detect HSP86.
PA3-012 has been successfully used in Western blot, immunohistochemistry, immunofluorescence, flow cytometry, and immunoprecipitation procedures. By Western blot, this antibody detects an ~84 kDa protein representing HSP84 from Hepa1 cell lysate. Immunofluorescence staining of HSP84 in Hepa1 cells with PA3-012 yields a pattern consistent with cytoplasmic staining. Immunoprecipitation experiments with this antibody suggest that HSP84 exists primarily as homodimers in Hepa 1 cells. Furthermore, the antibody is capable of precipitating HSP84 that is complexed with other proteins such as the aryl hydrocarbon (Ah) receptor.
The PA3-012 immunogen is a synthetic peptide corresponding to residues P(2) E E V H H G E E E V E(13) of mouse HSP84. The N-terminal regions of HSP84 and HSP86 show the largest difference in amino acid sequence.
Heat shock proteins (HSP) are expressed in response to various biological stresses, 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, aryl hydrocarbon (Ah) receptor, and some kinases.
Studies have shown that murine HSP90 exists as two forms, HSP84 and HSP86, coded by related but separate genes, with 86% amino acid sequence conservation. These forms are analogous to the two forms of human HSP90, HSP89 alpha and HSP89 beta. In an unstressed mouse fibroblast, the basal level of HSP84 is found to be double that of HSP86. However, after heat shock, HSP86 shows a greater increase. Studies also suggest that upon cellular differentiation, the level of HSP86, but not HSP84, decreases. HSP84 and HSP86, which may be subject to estrogenic regulation, have been found as components of the non-DNA binding form of mouse glucocorticoid receptor, but dissociated from the transformed DNA-binding form.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
The impact of disparate isolation methods for extracellular vesicles on downstream RNA profiling.
PA3-012 was used in western blot to assess the yield and purity of four different exosome isolation protocols
|Van Deun J,Mestdagh P,Sormunen R,Cocquyt V,Vermaelen K,Vandesompele J,Bracke M,De Wever O,Hendrix A||Journal of extracellular vesicles (3:null)||2014|
Vacuolar H+ ATPase expression and activity is required for Rab27B-dependent invasive growth and metastasis of breast cancer.
PA3-012 was used in western blot to study the mechanism by which vacuolar H(+)-ATPase regulates Rab27B-mediated invasion and metastases in estrogen receptor-positive breast cancer
|Hendrix A,Sormunen R,Westbroek W,Lambein K,Denys H,Sys G,Braems G,Van den Broecke R,Cocquyt V,Gespach C,Bracke M,De Wever O||International journal of cancer (133:843)||2013|
Heat shock protein 90¿ (HSP90¿), a substrate and chaperone of DNA-PK necessary for the apoptotic response.
PA3-012 was used in western blot to study the role of Hsp90 as both a substrate and chaperone of DNA-dependent protein kinase in the apoptotic response
|Solier S,Kohn KW,Scroggins B,Xu W,Trepel J,Neckers L,Pommier Y||Proceedings of the National Academy of Sciences of the United States of America (109:12866)||2012|
Effect of the secretory small GTPase Rab27B on breast cancer growth, invasion, and metastasis.
PA3-012 was used in western blot to investigate the role of secretory small GTPase Rab27B in breast cancer proliferation and metastasis
|Hendrix A,Maynard D,Pauwels P,Braems G,Denys H,Van den Broecke R,Lambert J,Van Belle S,Cocquyt V,Gespach C,Bracke M,Seabra MC,Gahl WA,De Wever O,Westbroek W||Journal of the National Cancer Institute (102:866)||2010|
Mutations that increase both Hsp90 ATPase activity in vitro and Hsp90 drug resistance in vivo.
PA3-012 was used in western blot to investigate the consequences of point mutations in heat shock protein 90 gene
|Zurawska A,Urbanski J,Matuliene J,Baraniak J,Klejman MP,Filipek S,Matulis D,Bieganowski P||Biochimica et biophysica acta (1803:575)||2010|
Proteomic identification of differentially expressed genes in mouse neural stem cells and neurons differentiated from embryonic stem cells in vitro.
PA3-012 was used in western blot to identify the differentially expressed proteins in neural stem cells and embryonic stem cells
|Akama K,Tatsuno R,Otsu M,Horikoshi T,Nakayama T,Nakamura M,Toda T,Inoue N||Biochimica et biophysica acta (1784:773)||2008|
Changes of endoplasmic reticulum chaperone complexes, redox state, and impaired protein disulfide reductase activity in misfolding alpha1-antitrypsin transgenic mice.
PA3-012 was used in western blot to investigate the changes of endoplasmic reticulum chaperone complexes, redox state, and impaired protein disulfide reductase activity in misfolding alpha1-antitrypsin transgenic mice
|Papp E,Száraz P,Korcsmáros T,Csermely P||FASEB journal : official publication of the Federation of American Societies for Experimental Biology (20:1018)||2006|
Comparative analysis of the ATP-binding sites of Hsp90 by nucleotide affinity cleavage: a distinct nucleotide specificity of the C-terminal ATP-binding site.
PA3-012 was used in western blot to study the specificity of the N-terminal and C-terminal Hsp90 nucleotide binding sites
|Soti C,Vermes A,Haystead TA,Csermely P||European journal of biochemistry (270:2421)||2003|
Subunit composition of the heteromeric cytosolic aryl hydrocarbon receptor complex.
PA3-012 was used in western blot to identify the subunits of AhR complex
|Chen HS,Perdew GH||The Journal of biological chemistry (269:27554)||1994|
Functional and cellular responses in a novel rodent model of anterior ischemic optic neuropathy.
PA3-012 was used in immunohistochemistry to evaluate a novel rodent anterior ischemic optic neuropathy model
|Bernstein SL,Guo Y,Kelman SE,Flower RW,Johnson MA||Investigative ophthalmology and visual science (44:4153)||2003|
Hsp and chaperone distribution during endochondral bone development in mouse embryo.
PA3-012 was used in immunohistochemistry to study the expression of heat shock proteins and chaperones during endochondral bone development in mouse
|Loones MT,Morange M||Cell stress and chaperones (3:237)||1998|
Function and regulation of heat shock factor 2 during mouse embryogenesis.
PA3-012 was used in immunocytochemistry to characterize heat shock factor 2 during mouse embryogenesis
|Rallu M,Loones M,Lallemand Y,Morimoto R,Morange M,Mezger V||Proceedings of the National Academy of Sciences of the United States of America (94:2392)||1997|