HSP90 alpha Antibody

Western Blot: HSP90 alpha Antibody [NB120-2928]

Western Blot: HSP90 alpha Antibody [NB120-2928] - Figure 1 shows a Western blot of mouse HSP84 and HSP86 using NB120-2928 and NB120-2927 respectively.

Western Blot: HSP90 alpha Antibody [NB120-2928]

Western Blot: HSP90 alpha Antibody [NB120-2928] - Analysis of epidermoid carcinoma (A431) cells using Heat Shock Protein 86 Polyclonal Antibody. Decreasing amounts of A431 whole cell lysates were probed with PA3-013 at a dilution of 1:2000 followed by an anti-rabbit IgG secondary antibody and SuperSignal West Pico Chemiluminescent Substrate. Lanes 1 and 2 are control samples of purified, recombinant Hsp86 (rHsp86) and Hsp84 (rHsp84), respectively. Data courtesy of the Innovators Program.

Western Blot: HSP90 alpha Antibody [NB120-2928]

Western Blot: HSP90 alpha Antibody [NB120-2928] - Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with a HSP86 polyclonal antibody at a dilution of 1:1000 overnight at 4C on a rocking platform, washed in TBS-0.1%Tween 20, and probed with a goat anti-rabbit IgG HRP secondary antibody at a dilution of 1:20,000 for at least 1 hour.

Western Blot: HSP90 alpha Antibody [NB120-2928]

Western Blot: HSP90 alpha Antibody [NB120-2928] - Analysis of 2-fold serial dilutions of HeLa cell lysate, starting at 10 ug, per well.

Immunohistochemistry-Paraffin: HSP90 alpha Antibody [NB120-2928]

Immunohistochemistry-Paraffin: HSP90 alpha Antibody [NB120-2928] - Both normal and cancer biopsies of deparaffinized Human kidney tissues.

Immunohistochemistry-Paraffin: HSP90 alpha Antibody [NB120-2928]

Immunohistochemistry-Paraffin: HSP90 alpha Antibody [NB120-2928] - Both normal and cancer biopsies of deparaffinized Human tonsil tissues.

Immunohistochemistry-Paraffin: HSP90 alpha Antibody [NB120-2928]

Immunohistochemistry-Paraffin: HSP90 alpha Antibody [NB120-2928] - Heat-induced epitope retrieval was performed using 10mM sodium citrate (pH 6.0) buffer for 20 mins at 95C. Following antigen retrieval, tissues blocked in 3% BSA in PBST for 30 mins at RT & then probed with Heat Shock Protein 86 (Hsp86) polyclonal antibody at 1:100 for 1 hr in a humidified chamber (right panel). As a -ve control, primary antibody eliminated from the staining procedure (left panel). Tissues washed with PBS/0.025% Tween-20 & endogenous peroxidase activity quenched with Peroxidase Suppressor for 30 mins at RT. Detection performed using an HRP-conjugated goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:250 followed by colorimetric detection using Metal Enhanced DAB Substrate Kit. Tissues were counterstained with hematoxylin & prepped for mouting. Images were taken on a Zeiss Axiovision microscope at 40X magnification (x1.6 Optovar).

Immunohistochemistry-Paraffin: HSP90 alpha Antibody [NB120-2928]

Immunohistochemistry-Paraffin: HSP90 alpha Antibody [NB120-2928] - Both normal and cancer biopsies of deparaffinized Human breast carcinoma tissues.

Immunohistochemistry: HSP90 alpha Antibody [NB120-2928]

HSP90-alpha-Antibody-Immunohistochemistry-NB120-2928-img0024.jpg

Immunohistochemistry: HSP90 alpha Antibody [NB120-2928]

HSP90-alpha-Antibody-Immunohistochemistry-NB120-2928-img0025.jpg

Immunoprecipitation: HSP90 alpha Antibody [NB120-2928]

Immunoprecipitation: HSP90 alpha Antibody [NB120-2928] - Analysis of Heat Shock Protein (HSP86) was performed on HeLa cells. Antigen-antibody complexes formed by incubating 500ug whole cell lysate with 2ug of HSP86 polyclonal antibody overnight on a rocking platform at 4C. The immune complexes were captured on 50ul Protein A/G Plus Agarose, washed extensively, and eluted with Lane Marker Reducing Sample Buffer. Samples were then resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with a HSP86 polyclonal antibody at a dilution of 1:1000 overnight rotating at 4C. The membrane was washed in TBST, and probed with Clean-Blot IP detection Reagent at a dilution of 1:1000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura.

Immunohistochemistry-Paraffin: HSP90 alpha Antibody [NB120-2928] -

Signaling consequences of STA-8666 treatment.A.-F. Left, Representative images and right, quantification of IHC or immunofluorescence for Ki-67 (A), cleaved caspase 3 (B), pS824-KAP1 (C), pT202/Y204-ERK1/2 (D), vimentin (light red) and DAPI (blue) (E) and HSP90 (F). Magnification: 20x. Scale bars: 75 μm, except for vimentin, where scale bar is 40 μm.

Western Blot: HSP90 alpha Antibody [NB120-2928] -

The “eHsp90 alpha > LRP-1 autocrine loop" only accounts for 50% of the constitutive motility of the tumour cells. (A) A schematic illustration of the “eHsp90 alpha > LRP-1 autocrine loop" that promotes cell motility. (B) Western blot shows 231-wt and two clones of 231-Hsp90 alpha -knockout (231-alpha -KO) cells (panel a). (C) Anti-Hsp90 alpha antibody blotting of secreted Hsp90 alpha in serum-free conditioned medium (CM, 10X) from 231-wt, 231-alpha -KO (panel c) or 231-LRP-1-KD (panel d) cells. (D) Western blot analysis shows the total Hsp90 alpha level in 231-wt or 231-LRP-1-KD cells under normoxia or hypoxia. (E) Western blot analysis shows the relative levels of LRP-1 in indicated cell lines. (F) CM from 231-wt, but not 231-alpha -KO, cells stimulated B16 melanoma cell migration (panels k and l vs. panel i), with human recombinant (hr) Hsp90 alpha protein (panel j) as positive control. (G) Computer-assisted quantitation of the B16 cell migration shown in panel F as Migration Index (%) (“Methods"). CM from 231-alpha -KO cells completely lost pro-motility activity (panel l). Asterisks indicate significance of the induced migration by indicated stimuli (panels j, k) versus the serum-free control (panel i). (H) The indicated cells were simultaneously compared for their motility under serum-fee conditions. Neither total alpha -KO (panels o and s) nor extracellular blockade of secreted Hsp90 alpha function by mAb1G7-D7 (panels p, t) re-produced the degree of inhibition of cell motility by LRP-1-KD (panels n and u), in reference to 231-wt cells (panels m and q). (L) Computer-assisted quantitation of the cell migration data with indicated panels. Asterisks indicate the significance of indicated cell motility over that of the LRP-1-KD cells. All experiments were repeated multiple times to reach the final conclusion. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35835845), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HSP90 alpha Antibody [NB120-2928] -

The “eHsp90 alpha > LRP-1 autocrine loop" only accounts for 50% of the constitutive motility of the tumour cells. (A) A schematic illustration of the “eHsp90 alpha > LRP-1 autocrine loop" that promotes cell motility. (B) Western blot shows 231-wt and two clones of 231-Hsp90 alpha -knockout (231-alpha -KO) cells (panel a). (C) Anti-Hsp90 alpha antibody blotting of secreted Hsp90 alpha in serum-free conditioned medium (CM, 10X) from 231-wt, 231-alpha -KO (panel c) or 231-LRP-1-KD (panel d) cells. (D) Western blot analysis shows the total Hsp90 alpha level in 231-wt or 231-LRP-1-KD cells under normoxia or hypoxia. (E) Western blot analysis shows the relative levels of LRP-1 in indicated cell lines. (F) CM from 231-wt, but not 231-alpha -KO, cells stimulated B16 melanoma cell migration (panels k and l vs. panel i), with human recombinant (hr) Hsp90 alpha protein (panel j) as positive control. (G) Computer-assisted quantitation of the B16 cell migration shown in panel F as Migration Index (%) (“Methods"). CM from 231-alpha -KO cells completely lost pro-motility activity (panel l). Asterisks indicate significance of the induced migration by indicated stimuli (panels j, k) versus the serum-free control (panel i). (H) The indicated cells were simultaneously compared for their motility under serum-fee conditions. Neither total alpha -KO (panels o and s) nor extracellular blockade of secreted Hsp90 alpha function by mAb1G7-D7 (panels p, t) re-produced the degree of inhibition of cell motility by LRP-1-KD (panels n and u), in reference to 231-wt cells (panels m and q). (L) Computer-assisted quantitation of the cell migration data with indicated panels. Asterisks indicate the significance of indicated cell motility over that of the LRP-1-KD cells. All experiments were repeated multiple times to reach the final conclusion. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35835845), licensed under a CC-BY license. Not internally tested by Novus Biologicals.