Anti-Hsp90 antibody [EPR3953]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 antibody [EPR3953] (AB109248)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung carcinoma tissue sections labeling Hsp90 with purified ab109248 at 1 : 50 dilution (19.42 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 antibody [EPR3953] (AB109248)

ab109248, at 1/100, staining Hsp90 in paraffin-embedded Human gastric adenocarcinoma tissue by Immunohistochemistry.

Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 antibody [EPR3953] (AB109248)

ab109248, at 1/100, staining Hsp90 in paraffin-embedded Human kidney tissue by Immunohistochemistry.

Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

Supplier Data

Western blot - Anti-Hsp90 antibody [EPR3953] (AB109248)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Hsp90 antibody [EPR3953] (ab109248) at 1/1000 dilution

Lane 1:

Western blot - Recombinant Human Hsp90 alpha protein (His tag) (<a href='/en-us/products/proteins-peptides/recombinant-human-hsp90-alpha-protein-his-tag-ab48801'>ab48801</a>) at 0.015 µg

Lane 2:

Western blot - Recombinant human Hsp90 beta protein (Active) (<a href='/en-us/products/proteins-peptides/recombinant-human-hsp90-beta-protein-active-ab80033'>ab80033</a>) at 0.015 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 85 kDa

Observed band size: 90 kDa

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Exposure time: 5s

• WB

Lab

Western blot - Anti-Hsp90 antibody [EPR3953] (AB109248)

All lanes:

Western blot - Anti-Hsp90 antibody [EPR3953] (ab109248) at 1/1000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 20 µg

Lane 2:

HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates at 20 µg

Lane 3:

RAW264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates at 20 µg

Lane 4:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates at 20 µg

Lane 5:

PC-12 (Rat adrenal gland pheochromocytoma ) whole cell lysates at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 85 kDa

Observed band size: 90 kDa

false

• WB

CiteAb

Western blot - Anti-Hsp90 antibody [EPR3953] (AB109248)

Hsp90 western blot using anti-Hsp90 antibody [EPR3953] ab109248. Publication image and figure legend from Guo, W., Yan, L., et al., 2014, PLoS One, PubMed 24465691.

ab109248 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab109248 please see the product overview.

MKT-077 potentiates 17-AAG-induced translocation of p53 to the nuclei in HCC cells.(A, B) Bel-7402 and HuH7 cells were treated with MKT-077, 17-AAG or a combination of both agents at the indicated concentrations for 24 hours. Cytoplasmic and nuclear fractions of Bel-7402 and HuH7 cells were separated, and subjected to Western blot analysis of p53 expression. α-tublin and Lamin B1 served as loading controls for cytoplasmic and nuclear fractions respectively. (C) Hep3B cells infected with Lenti-p53 (Hep3B-p53+/+) were treated with MKT-077, 17-AAG or MKT-077+17-AAG at indicated dosages for 24 hours, and immunofluorescent staining was performed to determine p53 subcellular localization. (D) Bel-7402 cells treated with MKT-077 or 17-AAG alone, or MKT-077+17-AAG for 24 hours were harvested and subjected to co-immunoprecipitation using GRP75-specific antibodies. Levels of p53, Akt and HSP90 were detected by Western blot analysis. (E) MKT-077 and 17-AAG synergistically enhanced mRNA levels of the p53 target genes p21 and PUMA, and reduced MDM2 mRNA levels. The mRNA levels of these genes were quantified using real-time RT-PCR and normalized to levels of GAPDH mRNA. * denotes p < 0.05 comparing 17-AAG to MKT-077+17-AAG.

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