重组Anti-p23抗体[EPR3846]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-p23 antibody [EPR3846] (AB92503)

Immunofluorescent staining of HeLa cells, using ab92503 at a dilution of 1/100.

• WB

Lab

Western blot - Anti-p23 antibody [EPR3846] (AB92503)

Lanes 1-4 : Merged signal (red and green). Green - ab92503 observed at 23 kDa. Red - loading control ab8245 observed at 36 kDa.

ab92503 Anti-p23 antibody [EPR3846] was shown to specifically react with p23 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266791 (knockout cell lysate ab258151) was used. Wild-type and p23 knockout samples were subjected to SDS-PAGE. ab92503 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-p23 antibody [EPR3846] (ab92503) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

PTGES3 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

Western blot - Human PTGES3 (p23) knockout HEK-293T cell line (<a href='/products/cell-lines/human-ptges3-p23-knockout-hek-293t-cell-line-ab266791'>ab266791</a>)

Lane 3:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Lane 4:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 19 kDa

Observed band size: 23 kDa

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• WB

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Western blot - Anti-p23 antibody [EPR3846] (AB92503)

All lanes:

Western blot - Anti-p23 antibody [EPR3846] (ab92503) at 1/10000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

Human spleen lysate at 10 µg

Lane 3:

Jurkat cell lysate at 10 µg

Lane 4:

HepG2 cell lysate at 10 µg

Secondary

All lanes:

HRP-labeled goat anti-rabbit at 1/2000 dilution

Predicted band size: 19 kDa

Observed band size: 23 kDa

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• WB

CiteAb

Western blot - Anti-p23 antibody [EPR3846] (AB92503)

p23 western blot using anti-p23 antibody [EPR3846] ab92503. Publication image and figure legend from He, Y., Peng, S., et al., 2016, Nat Commun, PubMed 27959342.

ab92503 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 ab92503 please see the product overview.

Interaction of AIL with p23 protein in vitro.(a) 22RV1 cells were treated with or without 1 μM AIL for 12 h. Anti-HSP90 IP was done and co-immunoprecipitated proteins were detected using indicated antibodies. (b) The interaction between p23 protein and AIL was measured by ProteOn XPR36. (c) Mapping AIL binding site on p23. (d) Top : expression level heatmap (log2 based) of AR1–651 induced genes which were inhibited by AIL. LNCaP cells were treated as described in Methods and the RNA samples of the indicated groups were sent for RNA-seq. First gene expression values (RPKM) were normalized (z-score transformed) across samples. Then the K-means clustering method was used to portion all genes into the clusters with Pearson correlation as the metric of distance. In the heat map, yellow means ‘higher' expression and blue means ‘lower' expression. Refseq IDs were converted into gene symbols listed in Supplementary Data 1. Bottom : Gene Ontology (GO) analysis of the AR1–651 target genes which inhibited by AIL. (e) Schematic illustrating the mechanism of downregulating AR protein level by AIL. When treating with AIL, the interaction of p23 and HSP90 is prevented and the interaction between AR and the molecular-chaperones is decreased, causing ubiquitination of AR. Then, AR is degraded by the proteasome, which reduces the expression of AR target genes and inhibits PCa growth and metastasis.

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