711744

antibody from Invitrogen Antibodies

Targeting: ERO1A ERO1-alpha, Ero1alpha, ERO1L

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

711744

Provider

Invitrogen Antibodies

Product name

ERO1L Recombinant Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Other

Description

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

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 SW480 (Lane 1), MCF-7 (Lane 2), MDA-MB-231 (Lane 3), T-47D (Lane 4), HCT 116 (Lane 5) and tissue extracts of mouse placenta (Lane 6) and mouse overy (Lane 7). The blots were probed with Anti-ERO1L Recombinant Rabbit Polyclonal Antibody (Product # 711744, 1 µg/mL). A 54 kDa band corresponding to Ero1 was observed across the cell lines and tissues tested. The blots were detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10% Bis-Tris gel (Product # NP0301BOX), 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

For immunofluorescence analysis, DU145 cells were fixed and permeabilized for detection of endogenous ERO1L using Anti- ERO1L Recombinant Rabbit Polyclonal Antibody (Product # 711744, 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 Ero1/endoplasmic oxidoreductin-1 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 Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating cytoplasmic localization of ERO1L. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.