Anti-FKBP52 antibody [EPR6618] - BSA and Azide free

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-FKBP52 antibody [EPR6618] - BSA and Azide free (AB240535)

This data was developed using ab129097, the same antibody clone in a different buffer formulation.

ab129097 staining FKBP52 in MCF-7 (human breast carcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% PFA and permeabilized with 0.1% triton X-100. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000.

DAPI was used as a nuclear counterstain and the negative control was PBS only.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FKBP52 antibody [EPR6618] - BSA and Azide free (AB240535)

This data was developed using ab129097, the same antibody clone in a different buffer formulation.

ab129097, at a 1/250 dilution, staining FKBP52 in paraffin embedded Human kidney tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FKBP52 antibody [EPR6618] - BSA and Azide free (AB240535)

This data was developed using ab129097, the same antibody clone in a different buffer formulation.

ab129097, at a 1/250 dilution, staining FKBP52 in paraffin embedded Human colon tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-FKBP52 antibody [EPR6618] - BSA and Azide free (AB240535)

This data was developed using ab129097, the same antibody clone in a different buffer formulation.

Western blot : Anti-FKBP52 antibody [EPR6618] ab129097 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 52 kDa in Wild-type HCT 116 cell lysates with no signal observed at this size in FKBP4 knockout HCT 116 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween 20 (TBS-T) before incubation with primary antibodies overnight at 4 C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-FKBP52 antibody [EPR6618] (<a href='/en-us/products/primary-antibodies/fkbp52-antibody-epr6618-ab129097'>ab129097</a>) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 at 20 µg

Lane 2:

Western blot - Human FKBP4 knockout HCT116 cell line (ab300946) at 20 µg

Lane 3:

LNCaP at 20 µg

Lane 4:

THP-1 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 52 kDa

Observed band size: 52 kDa

false

• WB

Unknown

Western blot - Anti-FKBP52 antibody [EPR6618] - BSA and Azide free (AB240535)

This data was developed using ab129097, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-FKBP52 antibody [EPR6618] (<a href='/en-us/products/primary-antibodies/fkbp52-antibody-epr6618-ab129097'>ab129097</a>) at 1/1000 dilution

Lane 1:

HeLa cell lysates at 10 µg

Lane 2:

Jurkat cell lysates at 10 µg

Lane 3:

K562 cell lysates at 10 µg

Lane 4:

MCF7 cell lysates at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 52 kDa

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-FKBP52 antibody [EPR6618] - BSA and Azide free (AB240535)

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.