Anti-CD3D antibody [EP4426] - Low endotoxin, Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CD3D antibody [EP4426] - Low endotoxin, Azide free (AB215040)

Immunocytochemistry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling CD3D with Purified ab109531 at 1 : 50 dilution (2.6 μg/ml). Cells were fixed in 100% Methanol 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 ab109531, the same antibody clone in a different buffer formulation.

• Flow Cyt

Lab

Flow Cytometry - Anti-CD3D antibody [EP4426] - Low endotoxin, Azide free (AB215040)

Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labelling CD3D with Purified ab109531 at 1 : 20 dilution (5μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) secondary antibody was used at 1 : 2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

This data was developed using ab109531, the same antibody clone in a different buffer formulation.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3D antibody [EP4426] - Low endotoxin, Azide free (AB215040)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue sections labeling CD3D with Purified ab109531 at 1 : 100 dilution (1.3 μg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 1 (pH 6.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1 : 0 dilution. PBS instead of the primary antibody was used as the negative control. M3

This data was developed using ab109531, the same antibody clone in a different buffer formulation.

• IP

Lab

Immunoprecipitation - Anti-CD3D antibody [EP4426] - Low endotoxin, Azide free (AB215040)

Purified ab109531 at 1/50 dilution (2μg) immunoprecipitating CD3D in Jurkat whole cell lysate.
Lane 1 (input) : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10μg
Lane 2 (+) : ab109531 + Jurkat whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab109531 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 : 23 kDa

This data was developed using ab109531, the same antibody clone in a different buffer formulation.

All lanes:

Immunoprecipitation - Anti-CD3D antibody [EP4426] (<a href='/en-us/products/primary-antibodies/cd3d-antibody-ep4426-ab109531'>ab109531</a>)

Predicted band size: 19 kDa

false

• WB

Supplier Data

Western blot - Anti-CD3D antibody [EP4426] - Low endotoxin, Azide free (AB215040)

This data was developed using ab109531, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-CD3D antibody [EP4426] (<a href='/en-us/products/primary-antibodies/cd3d-antibody-ep4426-ab109531'>ab109531</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

HuT-78 (Human Sezary syndrome cutaneous T lymphocyte) whole cell 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: 19 kDa

false