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Key Documents

03-102

Sigma-Aldrich

RIPAb+ HuR - RIP Validated Antibody and Primer Set

from rabbit

Synonym(s):

ELAV-like protein 1, Hu-antigen R

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.32

biological source

rabbit

Quality Level

clone

polyclonal

species reactivity

mouse, human, rat

manufacturer/tradename

RIPAb+
Upstate®

technique(s)

RIP: suitable
immunocytochemistry: suitable
immunohistochemistry: suitable
immunoprecipitation (IP): suitable
western blot: suitable

NCBI accession no.

UniProt accession no.

shipped in

dry ice

Gene Information

human ... ELAVL1(1994)

General description

RIPAb+ antibodies are evaluated using the RNA Binding Protein Immunoprecipitation (RIP) assay. Each RIPAb+ antibody set includes quantitative RT-PCR control primers (RIP Primers) to biologically validate your IP results by successfully coprecipitating a specific RNA target (where such a specific target is known). The qPCR protocol and primer sequences are provided, allowing researchers to validate RIP protocols when using the antibody in their experimental context. Each set also includes a negative control antibody to ensure specificity of the RIP reaction.
The RIPAb+ HuR set includes the HuR antibody, a negative control antibody (purified Rabbit IgG), and positive qPCR primers which amplify a 100 bp fragment of the human beta actin (ACTB) cDNA. The HuR and negative control antibodies are supplied in a scalable "per RIP" reaction size and can be used to functionally validate the precipitation HuR-associated RNAs.

Immunogen

Epitope: a.a. 1-13 of HuR

Application

Immunoprecipitation from RIP lysate:
RIP lysate from MCF7 cells (2 X 106 cell equivalents per IP) was subjected to immunoprecipitation using 0.5 μg of either a normal rabbit IgG or Anti-HuR antibody. Precipitated proteins were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-HuR (1.0 μg/mL).
Proteins were visualized using One-Step IP-Western kit (GenScript Cat. # L00231) (Please see figures).

Western Blot Analysis:
3T3/A31 lysate was resolved by electrophoresis, transferred to PVDF membranes and probed with HuR, hu (1:500 dilution). Proteins were visualized using a Donkey anti-Rabbit conjugated to HRP and visualized using a chemiluminescence detection system. (Please see figures).

Immunoprecipitation Analysis:
10 ug of this antibody immunoprecipitated HuR from 500 ug of 3T3/A31 RIPA lysate (Please see figures).

mmunocytochemistry Analysis:
Confocal IF analysis of HeLa, NIH/3T3 using anti-HuR (Red). Actin filaments have been labeled with AlexaFluor 488 -Phalloidin (Green). Nucleus is stained with DAPI (Blue) (Please see figures).
This RIPAb+ HuR -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

Quality

RNA binding protein Immunoprecipitation:
RIP Lysate prepared from HeLa cells (2 x 107 cell equivalents per IP) were subjected to immunoprecipitation using 5 µg of either a normal Rabbit IgG or Anti-HuR antibody and the Magna RIP Kit (Cat. # 17-700).
Successful immunoprecipitation of HuR-associated RNA was verified by qPCR using RIP Primers, ACTB (Please see figures).
Please refer to the Magna RIP® (Cat. # 17-700) or EZ-Magna RIP (Cat. # 17-701) protocol for experimental details.

Target description

35 kDa

Physical form

Anti-HuR (rabbit polyclonal). One vial containing 50 μg of purified rabbit polyclonal in 100 μL of buffer containing PBS in 0.05% sodium azide with 50% Glycerol. Liquid at -20°C.

Normal Rabbit IgG. One vial containing 125 μg purified rabbit IgG in 125 μL storage buffer containing 0.1% sodium azide. Store at -20°C.

RIP Primers, ACTB. One vial containing 75 μL of 5 μM of each primer specific for human beta actin (ACTB). Store at -20°C.
FOR: TTG TTA CAG GAA GTC CCT TGC C
REV: ATG CTA TCA CCT CCC CTG TGT G
Format: Purified

Analysis Note

Control
Included negative control rabbit IgG antibody and control primers specific for human beta actin (ATCB).

Legal Information

MAGNA RIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Storage Class Code

12 - Non Combustible Liquids

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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RNA-binding proteins play diverse roles in cancer, influencing various facets of the disease, including proliferation, apoptosis, angiogenesis, senescence, invasion, epithelial-mesenchymal transition (EMT), and metastasis. HuR, a known RBP, is recognized for stabilizing mRNAs containing AU-rich elements (AREs), although its complete
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Nature communications, 11(1), 45-45 (2020-01-04)
Unlike protein-coding genes, the majority of human long non-coding RNAs (lncRNAs) are considered non-conserved. Although lncRNAs have been shown to function in diverse pathophysiological processes in mice, it remains largely unknown whether human lncRNAs have such in vivo functions. Here

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