Anti-CD6/T12 antibody [EPR4057]

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-CD6/T12 antibody [EPR4057] (AB109217)

Intracellular Flow Cytometry analysis of HuT-78 (Human Sezary syndrome cutaneous T lymphocyte) cells, labeling CD6/T12 with Purified ab109217 at 1/20 dilution (10μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluorr^® 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).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD6/T12 antibody [EPR4057] (AB109217)

Unpurified ab109217 showing negative staining in Normal uterus tissue.

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-CD6/T12 antibody [EPR4057] (AB109217)

Unpurified ab109217, at a 1/100 dilution, staining CD6/T12 in paraffin embedded Human lymphocytes infiltrating colon tissue by Immunohistochemistry.

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

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-CD6/T12 antibody [EPR4057] (AB109217)

Immunocytochemistry/Immunofluorescence analysis of Human PBMC (Human primary peripheral blood mononuclear cell) cells labeling CD6/T12 with Purified ab109217 at 1 : 50 dilution (2.3 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 (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD6/T12 antibody [EPR4057] (AB109217)

Unpurified ab109217, at a 1/100 dilution, staining CD6/T12 in paraffin embedded Human tonsil 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-CD6/T12 antibody [EPR4057] (AB109217)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue sections labeling CD6/T12 with purified ab109217 at 1/1000 dilution (0.115 μg/mL). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IP

Unknown

Immunoprecipitation - Anti-CD6/T12 antibody [EPR4057] (AB109217)

Purified ab109217 at 1 : 20 dilution (0.6μg) immunoprecipitating CD6/T12 in HuT-78 whole cell lysate.
Lane 1 (input) : HuT-78 (Human Sezary syndrome cutaneous T lymphocyte) whole cell lysate 10μg
Lane 2 (+) : ab109217 + HuT-78 whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab109217 in HuT-78 whole cell lysate.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 105-130 kDa

All lanes:

Immunoprecipitation - Anti-CD6/T12 antibody [EPR4057] (ab109217)

Predicted band size: 72 kDa

false

• WB

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Western blot - Anti-CD6/T12 antibody [EPR4057] (AB109217)

The molecular weight of CD6 ranges from 105 to 130 kDa depending on its degree of phosphorylation (PMID : 17601777).

Blocking/Diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-CD6/T12 antibody [EPR4057] (ab109217) at 1/5000 dilution

All lanes:

HuT-78 (Human Sezary syndrome cutaneous T lymphocyte) whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 72 kDa

Observed band size: 105-130 kDa

false

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD6/T12 antibody [EPR4057] (AB109217)

Unpurified ab109217 showing negative staining in Normal brain tissue.

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-CD6/T12 antibody [EPR4057] (AB109217)

Unpurified ab109217 showing negative staining in Normal kidney tissue.

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-CD6/T12 antibody [EPR4057] (AB109217)

Unpurified ab109217 showing negative staining in Skeletal muscle tissue.

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

• WB

Unknown

Western blot - Anti-CD6/T12 antibody [EPR4057] (AB109217)

All lanes:

Western blot - Anti-CD6/T12 antibody [EPR4057] (ab109217) at 1/1000 dilution

Lane 1:

Jurkat lysates at 10 µg

Lane 2:

Human tonsil lysates at 10 µg

Lane 3:

PBMC lysates at 10 µg

Lane 4:

HuT-78 lysates at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 72 kDa

Observed band size: 90-130 kDa

false