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Western blotAntibody data
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- Western blot [3]
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- Product number
- 702709 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- ERO1L Recombinant Rabbit Monoclonal Antibody (11H3L5)
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- This antibody is predicted to react with Monkey, Pig, Bovine
- Antibody clone number
- 11H3L5
- Concentration
- 0.5 mg/mL
Submitted references Traumatic Optic Neuropathy Is Associated with Visual Impairment, Neurodegeneration, and Endoplasmic Reticulum Stress in Adolescent Mice.
Hetzer SM, Guilhaume-Correa F, Day D, Bedolla A, Evanson NK
Cells 2021 Apr 23;10(5)
Cells 2021 Apr 23;10(5)
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of ERO1 was achieved by transfecting MCF7 cells with ERO1 specific siRNA (Silencer® select Cat # s26883 and s26885). Western blot analysis (Fig a) was performed using Whole cell extract from the ERO1 knock down cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Anti-ERO1 Recombinant Rabbit Monoclonal Antibody (Product # 702709, 1-3 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). Densitometric analysis of this Western blot is shown in histogram (Fig b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to ERO1.
- 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), Hep G2 (Lane 2), HCT 116 (Lane 3), MCF-7 (Lane 4), HT-29 (Lane 5), MDA-MB-231 (Lane 6), HeLa (Lane 7), PANC-1 (Lane 8) and 3T3-L1 (Lane 9). The blots were probed with Anti-Ero1 Recombinant Rabbit Monoclonal Antibody (Product # 702709, 2.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:4000 dilution). A 54 kDa band corresponding to Ero1 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0322BOX), 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).
- 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), Hep G2 (Lane 2), HCT 116 (Lane 3), MCF-7 (Lane 4), HT-29 (Lane 5), MDA-MB-231 (Lane 6), HeLa (Lane 7), PANC-1 (Lane 8) and 3T3-L1 (Lane 9). The blots were probed with Anti-Ero1 Recombinant Rabbit Monoclonal Antibody (Product # 702709, 2.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:4000 dilution). A 54 kDa band corresponding to Ero1 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0322BOX), 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).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- For immunofluorescence analysis, SW480 cells were fixed and permeabilized for detection of endogenous ERO1 using Anti- ERO1 Recombinant Rabbit Monoclonal Antibody (Product # 702709, 5 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 (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 ERO1. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
