Anti-Brk/PTK6 antibody [EPR21051-96]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Brk/PTK6 antibody [EPR21051-96] (AB233392)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling Brk/PTK6 with ab233392 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic and nuclear staining in and membranous staining on human breast cancer cells is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Brk/PTK6 antibody [EPR21051-96] (AB233392)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized T-47D (human ductal breast epithelial tumor cell line) cell line labeling Brk/PTK6 with ab233392 at 1/600 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Brk/PTK6 antibody [EPR21051-96] (AB233392)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Brk/PTK6 with ab233392 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic and nuclear staining in surface epithelium of human colon (PMID : 10430081) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IP

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Immunoprecipitation - Anti-Brk/PTK6 antibody [EPR21051-96] (AB233392)

Brk/PTK6 was immunoprecipitated from 0.35 mg of T-47D (human ductal breast epithelial tumor cell line) whole cell lysate with ab233392 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab233392 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1 : T-47D whole cell lysate 10 μg (Input).

Lane 2 : ab233392 IP in T-47D whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab233392 in T-47D whole cell lysate.

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

Exposure time : 3 minutes.

IgG heavy chain : PMID : 18781181

All lanes:

Immunoprecipitation - Anti-Brk/PTK6 antibody [EPR21051-96] (ab233392)

Predicted band size: 52 kDa

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• WB

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Western blot - Anti-Brk/PTK6 antibody [EPR21051-96] (AB233392)

Exposure time : Lane 1 : 15 seconds; Lane 2-6 : 3 minutes.

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Brk/PTK6 antibody [EPR21051-96] (ab233392) at 1/1000 dilution

Lane 1:

T-47D (human ductal breast epithelial tumor cell line) whole cell lysate at 20 µg

Lane 2:

MCF7 (human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 3:

HT-29 (human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 4:

PC-3 (human prostate adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 5:

Human skin lysate at 20 µg

Lane 6:

Human breast cancer lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 52 kDa

Observed band size: 48 kDa

true