702406

antibody from Invitrogen Antibodies

Targeting: ESRRG ERR-gamma, ERRg, NR3B3

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

702406

Provider

Invitrogen Antibodies

Product name

ESRRG Recombinant Rabbit Monoclonal Antibody (4H7L11)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with Monkey, Sheep, Mouse

Antibody clone number

4H7L11

Concentration

0.5 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on Nuclear extracts (30 µg lysate) of SH-SY5Y (Lane 1), MCF 7 (Lane 2), MCF 7 (Treated with 20uM Retinoic acid in 1% FBS containing DMEM for 5 days) (Lane 3) and tissue extracts of Mouse Fetal Heart (Lane 4) and Rat Heart (Lane 5). The blots were probed with Anti-ESRRG Recombinant Rabbit Monoclonal Antibody (Product # 702406 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). Two isoforms of 51 kDa and 44 kDa band corresponding to ESRRG was observed across the cell lines and tissue 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 analysis of Estrogen-related receptor gamma was performed using 70% confluent log phase SH-SY5Y cells treated with Retinoic acid (20 µM, 5 days). The cells were fixed with 4% paraformaldehyde for 5 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for overnight at room temperature. The cells were labeled with ESRRG Recombinant Rabbit Monoclonal Antibody (4H7L11) (Product # 702406) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Nuclear localization. Panel e represents untreated cells with reduced signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.