Recombinant 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.

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). 

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

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.

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

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.

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.

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.

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.

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.

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.