Anti-OX40L/TNFSF4 antibody [EPR23155-317]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-OX40L/TNFSF4 antibody [EPR23155-317] (AB263910)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HUVEC (human umbilical vein endothelial cell) cells labelling CD252/OX40L with ab263910 at 1/100 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic and �staining in HUVEC cell line Negative control : HeLa (PMID : 1996093) is observed. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263910).

• Flow Cyt

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Flow Cytometry - Anti-OX40L/TNFSF4 antibody [EPR23155-317] (AB263910)

Flow cytometric analysis of HeLa (human cervix adenocarcinoma epithelial cell, Left) / HUVEC (human umbilical vein endothelial cell, Right) cells labelling CD252/OX40L with ab263910 at 0.002 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 anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263910).

• IP

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Immunoprecipitation - Anti-OX40L/TNFSF4 antibody [EPR23155-317] (AB263910)

OX40L/TNFSF4 was immunoprecipitated from 0.35 mg HUVEC (human umbilical vein endothelial cell) whole cell lysate 10µg with ab263910 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab263910 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : HUVEC (human umbilical vein endothelial cell) whole cell lysate 10µg
Lane 2 : ab263910 IP in HUVEC whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab263910 in HUVEC whole cell lysate

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

Exposure time : 20 seconds.

This antibody failed in WB test due to weak signals. But it can detect clear band after IP enrichment.

All lanes:

Immunoprecipitation - Anti-OX40L/TNFSF4 antibody [EPR23155-317] (ab263910)

Predicted band size: 21 kDa

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