Anti-PD1 antibody [29F.1A12] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PD1 antibody [29F.1A12] - BSA and Azide free (AB309361)

This data was developed using ab309360 the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Tween-20 permeabilized EL4 (mouse lymphoma T lymphocyte cell) cells labelling PD1 with ab309360 at 1/100 (9.8 ug/ml) dilution, followed by ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in EL4 cell line.Negative control : L-929 (mouse connective tissue fibroblast cell)Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab206369 Anti-beta Tubulin rabbit monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue). antibody only control : Secondary antibody is ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

• Flow Cyt

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Flow Cytometry - Anti-PD1 antibody [29F.1A12] - BSA and Azide free (AB309361)

This data was developed using ab309360 the same antibody clone in a different buffer formulation. Flow cytometric analysis of L-929 (mouse connective tissue fibroblast, Left) / EL4 (mouse lymphoma T lymphocyte, Right) cells labelling PD1 with ab309360 at 1/1000 dilution (0.1 ug)/Right compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat F(ab)2 Anti-Rat IgG Fc (Alexa Fluor® 488, ab150161) at 1/5000 dilution was used as the secondary antibody. Negative control : L-929 (PMID : 1396582). Gated on viable cells.

• WB

Supplier Data

Western blot - Anti-PD1 antibody [29F.1A12] - BSA and Azide free (AB309361)

This data was developed using ab309360, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST

The molecular weight observed is consistent with what has been described in the literature (PMID : 34635486).

This blot was developed using a high sensitivity ECL substrate.

All lanes:

Western blot - Anti-PD1 antibody [29F.1A12] (<a href='/en-us/products/primary-antibodies/pd1-antibody-29f1a12-ab309360'>ab309360</a>) at 1/1000 dilution

All lanes:

EL4.IL-2 (mouse lymphoma T lymphocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/100000 dilution

Observed band size: 50-55 kDa

false

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-PD1 antibody [29F.1A12] - BSA and Azide free (AB309361)

This data was developed using ab309360, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Negative control : L-929 (PMID : 1396582)

The molecular weight observed is consistent with what has been described in the literature (PMID : 34635486).

In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-PD1 antibody [29F.1A12] (<a href='/en-us/products/primary-antibodies/pd1-antibody-29f1a12-ab309360'>ab309360</a>) at 1/1000 dilution

Lane 1:

EL4 (mouse lymphoma T lymphocyte) whole cell lysate at 20 µg

Lane 2:

L-929 (mouse connective tissue fibroblast) whole cell lysate at 40 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/100000 dilution

Observed band size: 50-55 kDa

true

Exposure time: 37s