Anti-FKBP51 antibody [EPR6617] - BSA and Azide free

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FKBP51 antibody [EPR6617] - BSA and Azide free (AB242392)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat stomach tissue sections labeling FKBP51 with purified ab126715 at 1 : 250 dilution (1.156 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FKBP51 antibody [EPR6617] - BSA and Azide free (AB242392)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human prostatic hyperplasia tissue sections labeling FKBP51 with purified ab126715 at 1 : 250 dilution (1.156 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FKBP51 antibody [EPR6617] - BSA and Azide free (AB242392)

ab126715, at 1/50 dilution, stains FKBP51 in paraffin embedded human colon tissue by immunohistochemistry.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126715).

Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-FKBP51 antibody [EPR6617] - BSA and Azide free (AB242392)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling FKBP51 with purified ab126715 at 1/20 dilution (10 μg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-FKBP51 antibody [EPR6617] - BSA and Azide free (AB242392)

Immunocytochemistry/ Immunofluorescence analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling FKBP51 with purified ab126715 at 1 : 100 dilution (2.9 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1 : 200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IP

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Immunoprecipitation - Anti-FKBP51 antibody [EPR6617] - BSA and Azide free (AB242392)

ab126715 (purified) at 1 : 20 dilution (2μg) immunoprecipitating FKBP51 in Jurkat whole cell lysate.
Lane 1 (input) : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10μg
Lane 2 (+) : ab126715 & Jurkat whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab126715 in Jurkat whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1 : 1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-FKBP51 antibody [EPR6617] (<a href='/en-us/products/primary-antibodies/fkbp51-antibody-epr6617-ab126715'>ab126715</a>)

Predicted band size: 51 kDa

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• WB

Lab

Western blot - Anti-FKBP51 antibody [EPR6617] - BSA and Azide free (AB242392)

Lane 1 : Wild-type HAP1 whole cell lysate (20 μg)
Lane 2 : FKBP51 knockout HAP1 whole cell lysate (20 μg)
Lane 3 : HeLa whole cell lysate (20 μg)
Lane 4 : HepG2 whole cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab126715 observed at 51 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab126715 was shown to recognize FKBP51 in wild-type cells as signal was lost at the expected MW in FKBP51 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and FKBP51 knockout samples were subjected to SDS-PAGE. ab126715 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1,000 dilution and 1/20,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20,000 dilution for 1 hour at room temperature before imaging.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126715).

All lanes:

Western blot - Anti-FKBP51 antibody [EPR6617] (<a href='/en-us/products/primary-antibodies/fkbp51-antibody-epr6617-ab126715'>ab126715</a>)

Predicted band size: 51 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-FKBP51 antibody [EPR6617] - BSA and Azide free (AB242392)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.