Anti-PD-L2 antibody [CAL28]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-PD-L2 antibody [CAL28] (AB256386)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HDLM-2 (human Hodgkin lymphoma) cells labeling PD-L2 with ab256386 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG Isotype control (ab172730) (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-PD-L2 antibody [CAL28] (AB256386)

PD-L2 was immunoprecipitated from 0.35 mg of HDLM-2 (human Hodgkin lymphoma) whole cell lysate with ab256386 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab256386X at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.

Lane 1 : HDLM-2 whole cell lysate 10 μ (Input).
Lane 2 : ab256386 IP in HDML-2 whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab256386 in HDML-2 whole cell lysate.

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

All lanes:

Immunoprecipitation - Anti-PD-L2 antibody [CAL28] (ab256386)

Predicted band size: 31 kDa

Observed band size: 45-60 kDa

false

• WB

Supplier Data

Western blot - Anti-PD-L2 antibody [CAL28] (AB256386)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 10 seconds.

Negative control : Jurkat. (PMID : 27631416).

All lanes:

Western blot - Anti-PD-L2 antibody [CAL28] (ab256386) at 1/1000 dilution

Lane 1:

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

Lane 2:

Jurkat (human T cell leukemia cell line from peripheral blood) 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: 31 kDa

Observed band size: 45-60 kDa

false

• WB

Supplier Data

Western blot - Anti-PD-L2 antibody [CAL28] (AB256386)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 3 minutes.

All lanes:

Western blot - Anti-PD-L2 antibody [CAL28] (ab256386) at 1/1000 dilution

Lane 1:

MDA-MB-231 (human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 2:

U-2 OS (human bone osteosarcoma epithelial cell line) whole cell lysate at 20 µg

Lane 3:

MG-63 (human osteosarcoma fibroblast) whole cell lysate at 20 µg

Lane 4:

Saos-2 (human osteosarcoma cell line) whole cell lysate at 20 µg

Lane 5:

Human tonsil lysate at 20 µg

Lane 6:

Human lung lysate at 20 µg

Lane 7:

Human lung cancer 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: 31 kDa

Observed band size: 45-60 kDa

false