PA5-28749

antibody from Invitrogen Antibodies

Targeting: ESRRA ERR1, ERRa, ERRalpha, ESRL1, NR3B1

Western blot (Data presented on provider website)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Data presented on provider website)

Immunohistochemistry (Data presented on provider website)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-28749

Provider

Invitrogen Antibodies

Product name

ESRRA Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Recommended positive controls: 293T, A431, H1299, Mouse kidney, Rat kidney, A549. Predicted reactivity: Mouse (100%), Rat (100%), Dog (100%), Pig (100%), Bovine (100%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

1.32 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry analysis of ESRRA in MCF-7 cells. Samples were incubated in ESRRA polyclonal antibody (Product # PA5-28749) using a dilution of 1:500. Sample: MCF-7 cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: ERR alpha stained by ERR alpha antibody [N1], N-term. Red: alpha Tubulin, a cytoskeleton marker, stained by alpha Tubulin antibody [GT114] diluted at 1:1,000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of ESRRA was performed using 70% confluent log phase SK-BR-3 and MCF 10A cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with ESRRA Polyclonal Antibody (Product # PA5-28749) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing predominantly nuclear localization. Panel e represents MCF 10A cells having low expression of ESRRA. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Chromatin Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Cross-linked ChIP was performed with MCF-7 chromatin extract and 5 µg of either control rabbit IgG or ESRRA Polyclonal Antibody (Product # PA5-28749). The precipitated DNA was detected by PCR with primer set targeting to PS2 promoter or GAPDH promoter.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Chromatin Immunoprecipitation (ChIP) assay of endogenous ESRRA protein using Anti-ESRRA Antibody: ChIP was performed using Anti-ESRRA Rabbit Polyclonal Antibody (Product # PA5-28749) 5 µg, on sheared chromatin from etoposide treated MCF-7 cells using the MAGnify ChIP System kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to promoter of OGDH, IDH3A promoter and SAT2 satellite repeats. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

ERR alpha antibody immunoprecipitates ERR alpha protein in IP experiments. IP Sample: 293T whole cell lysate/extract A. 40 µg 293T whole cell lysate/extract B. Control with 2 µg of preimmune rabbit IgG C. Immunoprecipitation of ERR alpha protein by 2 µg of ERR alpha antibody (Product # PA5-28749) 7.5% SDS-PAGE The immunoprecipitated ERR alpha protein was detected by ERR alpha antibody (Product # PA5-28749) diluted at 1:1,000.