Antibodies, Proteins, Kits and Reagents for Life Science

351 792 ₸ Product size

100 µl 351 792 ₸ 10 µl 117 264 ₸

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Anti-FRA1 antibody [EPR23767-177] (ab252421)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR23767-177] to FRA1
Suitable for: Flow Cyt (Intra), WB, ICC, IP
Reacts with: Mouse, Human

Product name

Anti-FRA1 antibody [EPR23767-177]
See all FRA1 primary antibodies
Description

Rabbit monoclonal [EPR23767-177] to FRA1
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt (Intra)	Human
ICC	Human
IP	Human
WB	Mouse Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HeLa starved for 24 hours then treated with 100 ng/mL TPA whole cell lysate; NIH/3T3 starved for 2 hours then treated with 100 ng/mL TPA for 4 hours whole cell lysate; MDA-MB-231 whole cell lysate. ICC: HeLa cells. Flow cyt: HeLa cells starved for 24 hours then treated with 100 ng/ml TPA for 4 hours. IP: HeLa staved for 24 hours then treated with 100 ng/mL TPA for 4 hours.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR23767-177
Isotype

IgG
Research areas

Western blot - Anti-FRA1 antibody [EPR23767-177] (ab252421)

All lanes : Anti-FRA1 antibody [EPR23767-177] (ab252421) at 1/1000 dilution

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) staved for 24 hours, whole cell lysate
Lane 2 : HeLa staved for 24 hours then treated with 100ng /mL TPA (ab120297) for 4 hours, whole cell lysate
Lane 3 : NIH/3T3 (mouse embryonic fibroblast) staved for 2 hours, whole cell lysate
Lane 4 : NIH/3T3 staved for 2 hours then treated with 100 ng/mL TPA (ab120297) for 4 hours, whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution

Predicted band size: 29 kDa
Observed band size: 40 kDa
why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature (PMID:13679379).

The expression of FRA1 proteins is induced by TPA treatment (PMID: 13679379).

Exposure time: 3 minutes.
Immunocytochemistry - Anti-FRA1 antibody [EPR23767-177] (ab252421)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labelling FRA1 with ab252421 at 1/500 (0.918 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). The expression of FRA1 proteins is induced by TPA treatment (PMID: 13679379). Confocal image showing nuclear staining in HeLa cell line after starvation 24 hours, then treatment with Phorbol-12-myristate-13-acetate for 4h is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.
Flow Cytometry (Intracellular) - Anti-FRA1 antibody [EPR23767-177] (ab252421)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human cervix adenocarcinoma epithelial cell) starved for 24 hours then treated with 100 ng/ml TPA for 4 hours (Red) /Untreated control (Green) cells labelling FRA1 with ab252421 at 1/500 dilution (0.1ug) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunoprecipitation - Anti-FRA1 antibody [EPR23767-177] (ab252421)

FRA1 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) staved for 24 hours then treated with 100 ng/mL TPA for 4 hours, whole cell lysate with ab252421 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab252421 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) staved for 24 hours then treated with 100 ng/mL TPA for 4 hours, whole cell lysate 10 ug

Lane 2: ab252421 IP in HeLa staved for 24 hours then treated with 100 ng/mL TPA for 4 hours whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab252421 in HeLa staved for 24 hours then treated with 100 ng/mL TPA for 4 hours whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 15 seconds.
Western blot - Anti-FRA1 antibody [EPR23767-177] (ab252421)

Anti-FRA1 antibody [EPR23767-177] (ab252421) at 1/1000 dilution + MDA-MB-231 (human breast adenocarcinoma epithelial cell) whole cell lysate at 40 µg

Secondary
VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution

Predicted band size: 29 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST

Exposure time: 3 minutes.

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