Anti-Themis antibody [EPR7353] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Themis antibody [EPR7353] - BSA and Azide free (AB248177)

This data was developed using ab126771, the same antibody clone in a different buffer formulation.Intracellular Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling Themis with Purified ab248177 at 1/50 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).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Themis antibody [EPR7353] - BSA and Azide free (AB248177)

This data was developed using ab126771, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling Themis with Purified ab248177 at 1 : 50 dilution (10 ?g/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.

• IP

Lab

Immunoprecipitation - Anti-Themis antibody [EPR7353] - BSA and Azide free (AB248177)

This data was developed using ab126771, the same antibody clone in a different buffer formulation.
Purified ab126771 at 1/30 dilution (2μg) immunoprecipitating Themis in Jurkat whole cell lysate.
Lane 1 (input) : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10μg
Lane 2 (+) : ab126771 + Jurkat whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab126771 in Jurkat whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 73 kDa

All lanes:

Immunoprecipitation - Anti-Themis antibody [EPR7353] (<a href='/en-us/products/primary-antibodies/themis-antibody-epr7353-ab126771'>ab126771</a>)

Predicted band size: 73 kDa

false

• WB

Unknown

Western blot - Anti-Themis antibody [EPR7353] - BSA and Azide free (AB248177)

This data was developed using ab126771, the same antibody clone in a different buffer formulation.

The molecular weight observed is consistent with what has been described in the literature (PMID : 25535246 and PMID : 19597499).

Blocking Buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-Themis antibody [EPR7353] (<a href='/en-us/products/primary-antibodies/themis-antibody-epr7353-ab126771'>ab126771</a>) at 1/1000 dilution

Lane 1:

Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate at 20 µg

Lane 2:

MOLT-4 (Human lymphoblastic leukemia T lymphoblast) whole cell lysate at 20 µg

Lane 3:

Mouse thymus lysate at 20 µg

Lane 4:

Rat thymus lysate at 20 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: 73 kDa

Observed band size: 73 kDa

false