711309

antibody from Invitrogen Antibodies

Targeting: TNFRSF25 APO-3, DDR3, DR3, LARD, TNFRSF12, TR3, TRAMP, WSL-1, WSL-LR

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

711309

Provider

Invitrogen Antibodies

Product name

DR3 Recombinant Polyclonal Antibody (11HCLC)

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with Monkey, Cat, Pig, Sheep and Bovine.

Antibody clone number

11HCLC

Concentration

0.5 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) of K-562 (Lane 1), Jurkat (Lane 2), KARPAS 299 (Lane 3), THP-1 (Lane 4), PC-3 (Lane 5), SH-SY5Y (Lane 6), HeLa (Lane 7) and Caco-2 (Lane 8). The blots were probed with Anti-DR3/TNFRSF25 Recombinant Rabbit Polyclonal Antibody (Product # 711309, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 45kDa band along with 13 kDa isoform corresponding to DR3/TNFRSF25 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence was performed on fixed and permeabilized K562 cells for detection of endogenous DR3/TNFRSF25 using Anti- DR3/TNFRSF25 Recombinant Rabbit Polyclonal Antibody (Product # 711309, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of DR3/TNFRSF25 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating membrane localization of DR3/TNFRSF25. Panel e) represents control cells with no primary antibody to assess background.