RIPAb+ Anti-HuR, amino acids 3-19

货号： 03-102      产品名称： RIPAb+ Anti-HuR, amino acids 3-19   品牌： Millipore 规格： EA 3周到货 生化实验

RIPAb+ HuR – RIP Validated Antibody and Primer Set
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REFERENCES
Heat resistance of Bacillus spores when adhered to stainless steel and its relationship to spore hydrophobicity.
P Simmonds,B L Mossel,T Intaraphan,H C Deeth (2003) Journal of food protection.66
Species Reactivity Key Applications Host Format Antibody Type
H, M, R  RIP, IP, WB, ICC, IHC Rabbit Purified Polyclonal Antibody
Description:
RIPAb+ HuR – RIP Validated Antibody and Primer Set
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Trade Name:
Upstate (Millipore)
Product Overview:
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.
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Specificity:
HuR
Molecular Weight:
35 kDa
Epitope:
a.a. 1-13 of HuR
Immunogen:
KLH-conjugated, synthetic peptide corresponding to amino acids 1-13 of human HuR.
Background Information:
Hu-antigen R (HuB), ELAV-like protein 1, (ELAV1) is a transcription factor belonging to the ELAV family of RNP proteins. It binds avidly to the Au-rich element in cFos and interleukin-3 mRNAs.
Species Reactivity:
Human
Mouse
Rat
Application Notes:
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).
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Control:
Included negative control rabbit IgG antibody and control primers specific for human beta actin (ATCB).
Quality Assurance:
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 th -20°C