Anti-PD-L1 antibody [EPR20529]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PD-L1 antibody [EPR20529] (AB213480)

ab213480 staining PD-L1 in RAW264.7 from Mouse macrophage cell line transformed with Abelson murine leukemia virus by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 100% Methanol. Samples were incubated with ab213480 at 0.4 μg/ml. AlexaFluor^®488 Goat anti-Rabbit (ab150077) was used as the secondary antibody at 2 μg/ml. Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)(ab196889) was used as counterstain at 2.5 μg/ml. DAPI used as nuclear counterstain. Confocal image showing membranous staining on RAW 264.7 cells treated with IFN-γ (100 ng/ml) for 24 hours.

• IP

Supplier Data

Immunoprecipitation - Anti-PD-L1 antibody [EPR20529] (AB213480)

PD-L1 was immunoprecipitated from 0.35 mg of Mouse placenta lysate with ab213480 at 1/30 dilution.

Western blot was performed from the immunoprecipitate using ab213480 at 1/500 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1,000 dilution.

Lane 1 : Mouse placenta lysate, 10 μg (Input).

Lane 2 : ab213480 IP in mouse placenta lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab213480 in mouse placenta lysate.

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

Exposure time : 3 seconds.

All lanes:

Immunoprecipitation - Anti-PD-L1 antibody [EPR20529] (ab213480)

Predicted band size: 33 kDa

Observed band size: 55-60 kDa

false

• WB

Lab

Western blot - Anti-PD-L1 antibody [EPR20529] (AB213480)

This WB was performed by using YCA-R20529(BF)-111A H3L2 YR120921PJ, 1 : 5000 dilution. Working concentration : 0.402 μg/ml.

Blocking buffer and concentration : 5% NFDM/TBST

Diluting buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-PD-L1 antibody [EPR20529] (ab213480) at 1/5000 dilution

Lane 1:

Untreated RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 2:

RAW264.7 treated with 100 ng/ml IFN gamma for 48 h, 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 1/100000 dilution

Predicted band size: 33 kDa

Observed band size: 40-60 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-PD-L1 antibody [EPR20529] (AB213480)

Blocking/Dilution buffer : 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature. PMID : 15353579.

All lanes:

Western blot - Anti-PD-L1 antibody [EPR20529] (ab213480) at 1/1000 dilution

All lanes:

Mouse placenta lysate at 10 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: 33 kDa

Observed band size: 40-60 kDa

false

Exposure time: 15s

• WB

AbReview78135****

Western blot - Anti-PD-L1 antibody [EPR20529] (AB213480)

The mAb is specific for mouse mPDL1 and does not recognise hPDL1. As expected a specific 48kDa band is observed in RAW264.7 and MEFS cell extract after Interferon-gamma treatment. No bands are observed in RAW264.7 and MEFS WT and in MEFS KO with and without Interferon-gamma treatment.

All lanes:

Western blot - Anti-PD-L1 antibody [EPR20529] (ab213480) at 100 µg

Lane 1:

HEK mPDL1 cell lysate at 20 µg

Lane 2:

HEK hPDL1 cell lysate at 20 µg

Lane 3:

HEK CL7 at 20 µg

Lane 4:

RAW 264.7 Wild-type (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 100 µg

Lane 5:

RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate treated with Interferon-gamma at 100 µg

Lane 6:

MEF Wild-type (Mouse embryonic fibroblast cell line) whole cell lysate at 100 µg

Lane 7:

MEF (Mouse embryonic fibroblast cell line) whole cell lysate treated with Interferon-gamma at 100 µg

Lane 8:

MEF mPDL1 KO cell lysate at 100 µg

Lane 9:

MEF mPDL1 KO treated with Interferon-gamma at 100 µg

Predicted band size: 33 kDa

Observed band size: 48 kDa

false

Exposure time: 5min

This image is courtesy of an anonymous collaborator.