Immunofluorescent analysis of Heat Shock Protein 86 (HSP86, green) in HeLa cells and NIH3T3 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 with a HSP86 polyclonal antibody (Product # PA3-013), at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat-anti-rabbit IgG secondary antibody (Product # 35552) at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan at 20X magnification.
|Tested species reactivity||Human, Mouse, Non-human primate, Sheep, Rat|
|Published species reactivity||Rabbit, Non-human primate, Mouse, Human, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues P(2) E E T Q T Q D Q P M(12) of mouse HSP86.|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunofluorescence (IF)||1:50 - 1:200|
|Immunohistochemistry (Paraffin) (IHC (P))||5 µg/ml|
|Immunoprecipitation (IP)||2 µg|
|Western Blot (WB)||1:500 - 1:2000|
* 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.
PA3-013 detects heat shock protein 90 (HSP90) from human, monkey, mouse, sheep (adrenal) and rat tissues and cells. This antibody does not detect HSP84.
PA3-013 has been successfully used in Western blot, immunofluorescence, immunohistochemistry (paraffin) and immunoprecipitation procedures. By Western blot, this antibody detects an ~86 kDa protein representing HSP86 from HeLa cell lysate. Immunofluorescence staining of HSP86 in HeLa cells with PA3-013 yields a pattern consistent with cytoplasmic staining. Immunoprecipitation experiments with this antibody suggest that HSP86 exists primarily as homodimers in HeLa cells. Furthermore, the antibody is capable of precipitating HSP86 that is complexed with other proteins such as the aryl hydrocarbon (Ah) receptor.
The PA3-013 immunogen is a synthetic peptide corresponding to residues P(2) E E T Q T Q D Q P M(12) of mouse HSP86. 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, 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% homologous amino acid sequences. These forms are analogous to the two forms of human HSP90, HSP89 beta and HSP89 alpha. 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.
|Not Applicable||Not Cited||
LSD1 engages a corepressor complex for the activation of the estrogen receptor ¿ by estrogen and cAMP.
PA3-013 was used in western blot to assess activation of the estrogen receptor alpha by estrogen and cAMP and due to LSD1 engaging as a corepressor complex
|Bennesch MA,Segala G,Wider D,Picard D||Nucleic acids research (44:8655)||2016|
|Not Applicable||Not Cited||
Novobiocin Analogues That Inhibit the MAPK Pathway.
PA3-013 was used in western blot to analyze inhibitors of the MAPK pathway that are novobiocin analogues
|Hall JA,Seedarala S,Zhao H,Garg G,Ghosh S,Blagg BS||Journal of medicinal chemistry (59:925)||2016|
Integration-independent Transgenic Huntington Disease Fragment Mouse Models Reveal Distinct Phenotypes and Life Span in Vivo.
PA3-013 was used in western blot to assess transgenic mice expressing different forms of huntingtin protein for disease
|O'Brien R,DeGiacomo F,Holcomb J,Bonner A,Ring KL,Zhang N,Zafar K,Weiss A,Lager B,Schilling B,Gibson BW,Chen S,Kwak S,Ellerby LM||The Journal of biological chemistry (290:19287)||2015|
A C-terminal HSP90 inhibitor restores glucocorticoid sensitivity and relieves a mouse allograft model of Cushing disease.
PA3-013 was used in western blot to investigate how HSP90 affects the glucocorticoid receptor in corticotroph cells.
|Riebold M,Kozany C,Freiburger L,Sattler M,Buchfelder M,Hausch F,Stalla GK,Paez-Pereda M||Nature medicine (21:276)||2015|
Cruentaren A binds F1F0 ATP synthase to modulate the Hsp90 protein folding machinery.
PA3-013 was used in western blot to study the lack of induction of the pro-survival heat shock response following disruption of the interaction between F1F0 ATP synthase and Hsp90 by cruentaren A
|Hall JA,Kusuma BR,Brandt GE,Blagg BS||ACS chemical biology (9:976)||2014|
Vacuolar H+ ATPase expression and activity is required for Rab27B-dependent invasive growth and metastasis of breast cancer.
PA3-013 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|
Selective inhibition of HER2-positive breast cancer cells by the HIV protease inhibitor nelfinavir.
PA3-013 was used in western blot to study the ability of the HIV protease inhibitor nelfinavir to inhibit HER2-positive breast cancer cell growth via inhibition of Hsp90
|Shim JS,Rao R,Beebe K,Neckers L,Han I,Nahta R,Liu JO||Journal of the National Cancer Institute (104:1576)||2012|
A systematic protocol for the characterization of Hsp90 modulators.
PA3-013 was used in western blot to devise a method for the investigation of Hsp90 modulators
|Matts RL,Brandt GE,Lu Y,Dixit A,Mollapour M,Wang S,Donnelly AC,Neckers L,Verkhivker G,Blagg BS||Bioorganic and medicinal chemistry (19:684)||2011|
The molecular chaperone Hsp90¿ is required for meiotic progression of spermatocytes beyond pachytene in the mouse.
PA3-013 was used in western blot to study the role of Hsp90alpha in the meiotic progression of spermatocytes beyond pachytene in the mouse
|Grad I,Cederroth CR,Walicki J,Grey C,Barluenga S,Winssinger N,De Massy B,Nef S,Picard D||PloS one (5:null)||2010|
KU135, a novel novobiocin-derived C-terminal inhibitor of the 90-kDa heat shock protein, exerts potent antiproliferative effects in human leukemic cells.
PA3-013 was used in western blot to investigate the effect of a novel novobiocin-derived C-terminal Hsp90 inhibitor,KU135,on human leukemic cells
|Shelton SN,Shawgo ME,Matthews SB,Lu Y,Donnelly AC,Szabla K,Tanol M,Vielhauer GA,Rajewski RA,Matts RL,Blagg BS,Robertson JD||Molecular pharmacology (76:1314)||2009|
Pregnancy-enhanced endothelial nitric oxide synthase (eNOS) activation in uterine artery endothelial cells shows altered sensitivity to Ca2+, U0126, and wortmannin but not LY294002--evidence that pregnancy adaptation of eNOS activation occurs at multiple levels of cell signaling.
PA3-013 was used in western blot to investigate the roles of calcium and multiple kinases in the regulation of endothelial NO synthase activity.
|Sullivan JA,Grummer MA,Yi FX,Bird IM||Endocrinology (147:2442)||2006|
Domain-mediated dimerization of the Hsp90 cochaperones Harc and Cdc37.
PA3-013 was used in western blot to investigate the role of heat Shock Protein 90 cochaperones Harc and Cdc37 in protein folding and its mechanism
|Roiniotis J,Masendycz P,Ho S,Scholz GM||Biochemistry (44:6662)||2005|
Functional characterization of HUVEC-CS: Ca2+ signaling, ERK 1/2 activation, mitogenesis and vasodilator production.
PA3-013 was used in western blot to study Ca2+ signaling, ERK1/2 activation, mitogenesis and vasodilator production in an immortalized HUVEC subline
|Gifford SM,Grummer MA,Pierre SA,Austin JL,Zheng J,Bird IM||The Journal of endocrinology (182:485)||2004|
Purification of polyglutamine aggregates and identification of elongation factor-1alpha and heat shock protein 84 as aggregate-interacting proteins.
PA3-013 was used in western blot to investigate the roles of the EF-1alpha and HSP84 in the neurodegenerative process of polyglutamine diseases.
|Mitsui K,Nakayama H,Akagi T,Nekooki M,Ohtawa K,Takio K,Hashikawa T,Nukina N||The Journal of neuroscience : the official journal of the Society for Neuroscience (22:9267)||2002|
|Non-human primate||Not Cited||
Characterization of the phosphorylation status of the hepatitis B virus X-associated protein 2.
PA3-013 was used in western blot to indentify the phosphorylation status of the hepatitis B virus X-associated protein 2
|Dull AB,Carlson DB,Petrulis JR,Perdew GH||Archives of biochemistry and biophysics (406:209)||2002|
p50(cdc37) is a nonexclusive Hsp90 cohort which participates intimately in Hsp90-mediated folding of immature kinase molecules.
PA3-013 was used in western blot to investigate the role of p50(cdc37) in heat shock protein 90 molecular chaperone function
|Hartson SD,Irwin AD,Shao J,Scroggins BT,Volk L,Huang W,Matts RL||Biochemistry (39:7631)||2000|
Modular folding and evidence for phosphorylation-induced stabilization of an hsp90-dependent kinase.
PA3-013 was used in western blot to characterize a hsp90-dependent kinase and its regulation by phosphorylation
|Hartson SD,Ottinger EA,Huang W,Barany G,Burn P,Matts RL||The Journal of biological chemistry (273:8475)||1998|
Subunit composition of the heteromeric cytosolic aryl hydrocarbon receptor complex.
PA3-013 was used in western blot to identify the subunits of AhR complex
|Chen HS,Perdew GH||The Journal of biological chemistry (269:27554)||1994|
Subcellular heterogeneity of ryanodine receptor properties in ventricular myocytes with low T-tubule density.
PA3-013 was used in immunocytochemistry to characterize the ryanodine receptor in ventricular myocytes
|Biesmans L,Macquaide N,Heinzel FR,Bito V,Smith GL,Sipido KR||PloS one (6:null)||2011|
Transactivation of the epidermal growth factor receptor by heat shock protein 90 via Toll-like receptor 4 contributes to the migration of glioblastoma cells.
PA3-013 was used in immunocytochemistry to investigate the role of heat shock protein 90 in glioblastoma cell migration
|Thuringer D,Hammann A,Benikhlef N,Fourmaux E,Bouchot A,Wettstein G,Solary E,Garrido C||The Journal of biological chemistry (286:3418)||2011|
Function and regulation of heat shock factor 2 during mouse embryogenesis.
PA3-013 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|
Hsp and chaperone distribution during endochondral bone development in mouse embryo.
PA3-013 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|