Anti-PD-L1 antibody [RM1012] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [RM1012] - BSA and Azide free (AB283867)

This data was developed using ab282458, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labelling PD-L1 with ab282458 at 1/500 (0.876 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human breast cancer. The section was incubated with ab282458 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [RM1012] - BSA and Azide free (AB283867)

This data was developed using ab282458, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labelling PD-L1 with ab282458 at 1/500 (0.876 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human tonsil. The section was incubated with ab282458 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [RM1012] - BSA and Azide free (AB283867)

Immunohistochemical analysis of paraffin-embedded Human placenta tissue labelling PD-L1 with ab282458 at 1/500 (0.876 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human placenta. The section was incubated with ab282458 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [RM1012] - BSA and Azide free (AB283867)

This data was developed using ab282458, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human lung cancer tissue labelling PD-L1 with ab282458 at 1/500 (0.876 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human lung cancer. The section was incubated with ab282458 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• WB

Supplier Data

Western blot - Anti-PD-L1 antibody [RM1012] - BSA and Azide free (AB283867)

This data was developed using ab282458, the same antibody clone in a different buffer formulation.

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

Low expression control : MCF7 (PMID : 28184013, 31741201).

All lanes:

Western blot - Anti-PD-L1 antibody [RM1012] - BSA and Azide free (ab283867) at 1/1000 dilution

Lane 1:

NCI-H1975 (Human adenocarcinoma lung epithelial cell) whole cell lysate at 20 µg

Lane 2:

MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

Human placenta 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

• WB

Supplier Data

Western blot - Anti-PD-L1 antibody [RM1012] - BSA and Azide free (AB283867)

This data was developed using ab282458, the same antibody clone in a different buffer formulation.

PD-L1 was immunoprecipitated from 0.35 mg NCI-H1975 (Human adenocarcinoma lung epithelial cell) whole cell lysate with ab282458 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab282458 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : NCI-H1975 (Human adenocarcinoma lung epithelial cell) whole cell lysate 10 ug

Lane 2 : ab282458 IP in NCI-H1975 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab282458 in NCI-H1975 whole cell lysate

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

Exposure time : 32 seconds

All lanes:

Western blot - Anti-PD-L1 antibody [RM1012] - BSA and Azide free (ab283867)

Predicted band size: 33 kDa

Observed band size: 40-60 kDa

false

• ICC

Supplier Data

Immunocytochemistry - Anti-PD-L1 antibody [RM1012] - BSA and Azide free (AB283867)

This data was developed using ab282458, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized CHO-PDL1 cells labelling PD-L1 with ab282458 at 1/1000 (0.438 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in CHO-PDL1 cells and no staining in CHO-S cells.

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

• Flow Cyt

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Flow Cytometry - Anti-PD-L1 antibody [RM1012] - BSA and Azide free (AB283867)

This data was developed using ab282458, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of CHO-S (Chinese hamster ovary epithelial cell (Blue)) / CHO-PDL1 (PD-L1 stably expessed Chinese hamster ovary epithelial cell) (Red)) cells labelling PD-L1 with ab282458 at 1/50 dilution (1ug). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.

• I-ELISA

Supplier Data

Indirect ELISA - Anti-PD-L1 antibody [RM1012] - BSA and Azide free (AB283867)

This data was developed using ab282458, the same antibody clone in a different buffer formulation.

ELISA using ab282458 at varying antibody concentrations and antigen concentration at 1000 ng/ml. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) (1/2500) was used as the secondary antibody.