Anti-CD6/T12 antibody [EPR4057] - BSA and Azide free

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CD6/T12 antibody [EPR4057] - BSA and Azide free (AB247795)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109217).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD6/T12 antibody [EPR4057] - BSA and Azide free (AB247795)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109217).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-CD6/T12 antibody [EPR4057] - BSA and Azide free (AB247795)

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 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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109217).

• IP

Unknown

Immunoprecipitation - Anti-CD6/T12 antibody [EPR4057] - BSA and Azide free (AB247795)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109217).

All lanes:

Immunoprecipitation - Anti-CD6/T12 antibody [EPR4057] (<a href='/en-us/products/primary-antibodies/cd6-t12-antibody-epr4057-ab109217'>ab109217</a>)

Predicted band size: 72 kDa

false

• WB

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Western blot - Anti-CD6/T12 antibody [EPR4057] - BSA and Azide free (AB247795)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109217).

All lanes:

Western blot - Anti-CD6/T12 antibody [EPR4057] (<a href='/en-us/products/primary-antibodies/cd6-t12-antibody-epr4057-ab109217'>ab109217</a>) 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