Recombinant Anti-FKBP52 antibody [EPR21125] (ab230951)

Lanes 1-2 : Anti-FKBP52 antibody [EPR21125] (ab230951) at 1/5000 dilution
Lanes 3-4 : Anti-FKBP52 antibody [EPR21125] (ab230951) at 1/1000 dilution

Lane 1 : K562 (human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate at 20 µg
Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 3 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 4 : Human fetal brain lysate at 10 µg

Secondary
Lanes 1-3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lane 4 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

Predicted band size: 52 kDa
Observed band size: 52 kDa

Blocking/Diluting buffer: 5% NFDM/TBST.

Exposure times: Lane 1-2: 48 seconds; Lane 3: 30 seconds; Lane 4: 3 minutes.

The molecular mass observed is consistent with what has been described in the literature (PMID: 26065228).

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling FKBP52 with ab230951 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and weakly nuclear staining in HeLa cell line. 

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red). The nuclear counter stain is DAPI (blue). 

Secondary antibody only control: Used PBS instead of the primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

FKBP52 was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab230951 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab230951 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1: HeLa whole cell lysate 10 µg (Input). 

Lane 2: ab230951 IP in HeLa whole cell lysate. 

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab230951 in HeLa whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 30 seconds.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF7 (human breast adenocarcinoma cell line) cells labeling FKBP52 with ab230951 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and weakly nuclear staining in MCF7 cell line.

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red). The nuclear counter stain is DAPI (blue).

Secondary antibody only control: Used PBS instead of the primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling FKBP52 with ab230951 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^� 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.