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03-102
Sigma-AldrichRIPAb+ HuR - RIP Validated Antibody and Primer Set
This RIPAb+ HuR -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
More>>This RIPAb+ HuR -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers. Less<<
SDB (Sicherheitsdatenblätter), Analysenzertifikate und Qualitätszertifikate, Dossiers, Broschüren und andere verfügbare Dokumente.
RIPAb+ HuR - RIP Validated Antibody and Primer Set
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.
Alternate Names
ELAV-like protein 1
Hu-antigen R
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.
References
Product Information
Format
Purified
Control
Included negative control rabbit IgG antibody and control primers specific for human beta actin (ATCB).
Presentation
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
This RIPAb+ HuR -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Key Applications
RNA Binding Protein Immunoprecipitation (RIP)
Immunoprecipitation
Western Blotting
Immunocytochemistry
Immunohistochemistry
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).
Biological Information
Immunogen
KLH-conjugated, synthetic peptide corresponding to amino acids 1-13 of human HuR.
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.
FUNCTION: SwissProt: Q15717 # Binds avidly to the AU-rich element in FOS and IL3/interleukin-3 mRNAs. In the case of the FOS AU-rich element, HUR binds to a core element of 27 nucleotides that contain AUUUA, AUUUUA, and AUUUUUA motifs. SIZE: 326 amino acids; 36092 Da SUBUNIT: Interacts with ANP32A. TISSUE SPECIFICITY: Ubiquitous. PTM: Methylated at Arg-217 by CARM1 in macrophages in response to LPS challenge. SIMILARITY: SwissProt: Q15717 ## Belongs to the RRM elav family. & Contains 3 RRM (RNA recognition motif) domains.
Molecular Weight
35 kDa
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
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 the Magna RIP™ (Cat. # 17-700) or EZ-Magna RIP™ (Cat. # 17-701) protocol for experimental details.
Usage Statement
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage Conditions
Stable for 1 year at -20°C from date of receipt. Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.
Packaging Information
Material Size
10 assays
Material Package
10 assays per set. Recommended use: ~5 μg of antibody per RIP (dependent upon biological context).
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Bestellnummer
GTIN
03-102
04053252583209
Documentation
RIPAb+ HuR - RIP Validated Antibody and Primer Set SDB
The recognition of stress-induced ligands by the activating receptor NKG2D expressed on cytotoxic lymphocytes is crucial for the prevention and containment of various diseases and is also one of the best-studied examples of how danger is sensed by the immune system. Still, however, the mechanisms leading to the expression of the NKG2D ligands are far from being completely understood. Here, we use an unbiased and systematic RNA pull-down approach combined with mass spectrometry to identify six RNA-binding proteins (RBPs) that bind and regulate the expression of MICB, one of the major stress-induced ligands of NKG2D. We further demonstrate that at least two of the identified RBPs function during genotoxic stress. Our data provide insights into stress recognition and hopefully open new therapeutic venues.
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 Journal of food protection
66
2003
Twenty-one strains of Bacillus (10 B. stearothermophilus, 3 B. cereus, and 8 B. licheniformis strains) were assayed for spore surface hydrophobicity on the basis of three measures: contact angle measurement (CAM), microbial adhesion to hydrocarbons (MATH), and hydrophobic interaction chromatography (HIC). On the basis of the spore surface characteristics obtained from these assays, along with data on the heat resistance of these spores in water, eight strains of Bacillus (three B. stearothermophilus, three B. cereus, and two B. licheniformis strains) either suspended in water or adhering to stainless steel were exposed to sublethal heat treatments at 90 to 110 degrees C to determine heat resistance (D-value). Significant increases in heat resistance (ranging from 3 to 400%) were observed for the eight strains adhering to stainless steel. No significant correlation was found between these heat resistance increases and spore surface characteristics as determined by the three hydrophobicity assays. There was a significant positive correlation between the hydrophobicity data obtained by the MATH assay and those obtained by the HIC assay, but these data did not correlate with those obtained by the CAM assay.
