Invitrogen

HuR Monoclonal Antibody (3A2)

Advanced Verification

This Antibody was verified by Relative expression to ensure that the antibody binds to the antigen stated.

1 Reference

View all (22) HuR antibodies

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Datasheet

Protocols

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HuR Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of HuR was performed using 70% confluent log phase HeLa cells treated with Actinomycin D (1uM for 19hrs). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with HuR Monoclonal Antibody (3A2) (Product # MA1-167) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 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 translocation of HuR in to the cytoplasm. Panel e represents the control cells showing exclusive nuclear staining. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

HuR Antibody in Western Blot (WB) — Western blot analysis of HuR (3A2) was performed by loading ~6-20 µg of the indicated whole cell lysates per well, and 5 µL of PageRuler Plus Prestained Protein Ladder (Product # 26619) onto a 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane using the G2 Fast Blotter (Product # 62288), and blocked with 5% milk in TBST for 1 hour at room temperature. HuR was detected at ~36 kD in samples using a HuR (3A2) monoclonal antibody (Product # MA1-167) at a dilution of 1:250 overnight at 4C on a rocking platform, followed by a goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:10,000 for at least 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico substrate (Product # 34080).

HuR Antibody in Immunoprecipitation (IP) — Immunoprecipitation of HuR was performed using MCF-7 cells. Antigen-antibody complexes were formed by incubating 500 µg of MCF-7 whole cell lysates with 2 µg of HuR (3A2) monoclonal antibody (Product # MA1-167) overnight on a rocking platform at 4C. The immune complexes were captured on 100 µL Protein A/G Agarose (Product # 20421), washed extensively, and eluted with 5X Lane Marker Reducing Sample Buffer (Product # 39000). The IP eluate (right lane) and 20 µg of MCF-7 lysate (left lane) were resolved on a 4-20% Tris-Glycine polyacrylamide gel, transferred to a nitrocellulose membrane, and blocked with 5% Milk in TBST for at least 1 hour. The membrane was probed with a HuR (3A2) monoclonal antibody monoclonal antibody (Product # MA1-167) at a dilution of 1:250 overnight rotating at 4C, washed in TBST, and probed with goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:10,000 for at least 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080).

HuR Antibody in RNA Immunoprecipitation (RIP) — RNA immunoprecipitation (RIP) western of ELAVL1 was performed on K562 cells. Antigen-antibody complexes were formed by incubating approximately 500 µg whole cell lysate with 5 µg of monoclonal ELAVL1 antibody (Product # MA1-167) rotating 60 min at RT. The immune complexes were captured on 625 µg of Dynabeads M-280 sheep anti-mouse IgG (Product # 11202D), washed extensively, and eluted in NuPAGE LDS sample buffer (Product # NP0007). Samples were resolved onto a 4-12% Bis-Tris protein gel (Product # NP0335BOX). Lanes 1 and 3 are input and lanes 2 and 4 are IP. Proteins were transferred to nitrocellulose membrane (Product # IB23001) and blocked in 5% milk. ELAVL1 was detected at approximately 36 kDa using a monoclonal ELAVL1 antibody (Product # MA1-167) at a dilution of 1:2000, followed by incubation with anti-mouse secondary antibody. Chemiluminescent detection was performed using ECL Western Blotting Substrate (Product # 32106). Data courtesy of the Yeo lab as part of the ENCODE project (www.encodeproject.org).

HuR Antibody — Antibody specificity was demonstrated by detection of binding of the target protein to specific RNA. RNA Immunoprecipitation (RIP) was performed using Anti-HuR Monoclonal Antibody (Product # MA1-167) using primer pairs over ACTB mRNA (positive) and 18S rRNA (inactive). {RE}

Product Details

MA1-167

Applications	Tested Dilution	Publications
Western Blot (WB)	1:125-1:500	-
Immunocytochemistry (ICC/IF)	1:25-1:100	View 1 publication 1 publication
Immunoprecipitation (IP)	2 µg/10^6 cells	-
RNA Immunoprecipitation (RIP)	2 µg/10^6 cells	-

Product Specifications
Species Reactivity	Human, Mouse, Rat
Published species	Rat
Host/Isotype	Mouse / IgG1
Class	Monoclonal
Type	Antibody
Clone	3A2
Immunogen	Recombinant human HuR protein
Conjugate	Unconjugated
Form	Liquid
Concentration	0.5 mg/mL
Purification	Protein A
Storage buffer	PBS, pH 7.2-7.4, with 1mg/mL BSA, 30% glycerol
Contains	0.05% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_2536870

Target Information

The protein encoded by this gene is a member of the ELAVL protein family. This encoded protein contains 3 RNA-binding domains and binds cis-acting AU-rich elements. It destabilizes mRNAs and thereby regulates gene expression.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

RNA-binding protein HuR regulates translation of vitamin D receptor modulating rapid epithelial restitution after wounding.

American journal of physiology. Cell physiology

Zhang Y,Cai JZ,Xiao L,Chung HK,Ma XX,Chen LL,Rao JN,Wang JY

MA1-167 was used in Immunocytochemistry to indicate that HuR enhances VDR translation by directly interacting with its mRNA via 3'-UTR and that induced VDR by HuR is crucial for rapid intestinal epithelial restitution after wounding.

Wed Jul 01 00:00:00 EDT 2020

Bioinformatics

Protein Aliases: ELAV (embryonic lethal, abnormal vision, Drosophila)-like 1 (Hu antigen R); ELAV like 1; Elav-like generic protein; ELAV-like protein 1; embryonic lethal, abnormal vision, drosophila, homolog-like 1; Hu antigen R; HU-antigen A; Hu-antigen R; Human antigen R; HuR; MelG

Gene Aliases: 2410055N02Rik; ELAV1; ELAVL1; Elra; Hua; HUR; MelG; W91709

UniProt ID: (Human) Q15717, (Mouse) P70372, (Rat) B5DF91

Entrez Gene ID: (Human) 1994, (Mouse) 15568, (Rat) 363854

Molecular Function: translation initiation factor