Anti-PD-L2 antibody [EPR25200-50] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L2 antibody [EPR25200-50] - BSA and Azide free (AB288308)

This data was developed using ab288298, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human endometrial cancer tissue labelling PD-L2 with ab288298 at 1/1000 (0.546 ug/ml) followed by a ready to use LeicaDS9800 (Bond™, Polymer Refine Detection) was used. Positive staining on human endometrial cancer (PMID : 27446374). The section was incubated with ab288298 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) was used.

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L2 antibody [EPR25200-50] - BSA and Azide free (AB288308)

This data was developed using ab288298, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Panel A HDLM-2 Panel B Jurkat labelling PD-L2 with ab288298 at 1/1000 (0.546 ug/ml) followed by a ready to use LeicaDS9800 (Bond™, Polymer Refine Detection) was used. Positive cell line HDLM-2 (Panel A) was positive and negative cell line Jurkat (Panel B) was negative.The section was incubated with ab288298 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 Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L2 antibody [EPR25200-50] - BSA and Azide free (AB288308)

This data was developed using ab288298, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labelling PD-L2 with ab288298 at 1/1000 (0.546 ug/ml) followed by a ready to use LeicaDS9800 (Bond™, Polymer Refine Detection) was used. Positive staining on human breast cancer. The section was incubated with ab288298 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 Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L2 antibody [EPR25200-50] - BSA and Azide free (AB288308)

This data was developed using ab288298, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labelling PD-L2 with ab288298 at 1/1000 (0.546 ug/ml) followed by a ready to use LeicaDS9800 (Bond™, Polymer Refine Detection) was used. Positive staining is found in the germinal centre of human tonsil. The section was incubated with ab288298 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 Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IP

Supplier Data

Immunoprecipitation - Anti-PD-L2 antibody [EPR25200-50] - BSA and Azide free (AB288308)

This data was developed using ab288298, the same antibody clone in a different buffer formulation.

PD-L2 was immunoprecipitated from 0.35 mg HDLM-2 (human Hodgkin lymphoma) whole cell lysate with ab288298 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab288298 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HDLM-2 (human Hodgkin lymphoma) whole cell lysate 10 ug

Lane 2 : ab288298 IP in HDLM-2 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab288298 in HDLM-2 whole cell lysate

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

Exposure time : 92 seconds

All lanes:

Immunoprecipitation - Anti-PD-L2 antibody [EPR25200-50] (<a href='/en-us/products/primary-antibodies/pd-l2-antibody-epr25200-50-ab288298'>ab288298</a>)

Predicted band size: 31 kDa

Observed band size: 45-60 kDa

false

• WB

Lab

Western blot - Anti-PD-L2 antibody [EPR25200-50] - BSA and Azide free (AB288308)

This data was developed using ab288298, the same antibody clone in a different buffer formulation.

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

Negative control : Jurkat (PMID : 27631416).

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-PD-L2 antibody [EPR25200-50] (<a href='/en-us/products/primary-antibodies/pd-l2-antibody-epr25200-50-ab288298'>ab288298</a>) at 1/1000 dilution

Lane 1:

HDLM-2 (human Hodgkin lymphoma) whole cell lysate at 20 µg

Lane 2:

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

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 31 kDa

Observed band size: 45-60 kDa

false

Exposure time: 125s