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03-242 Sigma-Aldrich

RIPAb+ PUM1 Antibody

from rabbit, purified by affinity chromatography

Synonym: pumilio (Drosophila) homolog 1, pumilio 1, pumilio homolog 1 (Drosophila)

  •  eCl@ss 32160702

  •  NACRES NA.32

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Properties

Related Categories Antibodies, Antibody Kits, Immunoprecipitation Kits More...
clone   polyclonal
biological source   rabbit
application(s)   RIP: suitable
  immunocytochemistry: suitable
  western blot: suitable
species reactivity   primate, mouse, mouse, bovine, rat, human, rat, rhesus macaque, human, chimpanzee
species reactivity (predicted by homology)   primate (based on 100% sequence homology), bovine (based on 100% sequence homology), rhesus monkey (based on 100% sequence homology), chimpanzee (based on 100% sequence homology)
shipped in   dry ice
Quality Level   100
purified by   affinity chromatography
mfr. no.   RIPAb+
  Upstate®
NCBI accession no.   NP_001018494
UniProt accession no.   Q14671
Gene Information   human ... PUM1(9698)

Description

General description

RIPAb+ antibodies are evaluated using the RNA Binding Protein Immunoprecipitation (RIP) assay. Each RIPAb+ antibody set includes a negative control antibody to ensure specificity of the RIP reaction and is verified for the co-immunoprecipitation of RNA associated specifically with the immunoprecipitated RNA-binding protein of interest. Where appropriate, the RIPAb+ set also includes quantitative RT-PCR control primers (RIP Primers) to biologically validate your IP results by successfully co-precipitating the specific RNA targets, such as messenger RNAs. The qPCR protocol and primer sequences are provided, allowing researchers to validate RIP protocols when using the antibody in their experimental context. If a target specific assay is not provided, the RIPAb+ kit is validated using an automated microfluidics-based assay by enrichment of detectable RNA over control immunoprecipitation.
PUM1 has been shown to bind an mRNA sequence known as Nanos response element. It is found in heart, brain, muscle, kidney, stomach and intestine. PUM proteins play an active role in the sustainability of mitotic stem cell proliferation and the regulation of cell development and differentiation.

Within the mammalian genome are two genes belonging to a family of housekeeping genes known as PUF. These genes are known as PUM1 and PUM2. PUF proteins are characterized by having a C-terminal RNA-binding domain called the Pumilio homolog domain, which affects the stability of mRNA and translation by binding the 3’-UTR of mRNA targets. PUM plays an active role in the sustainability of mitotic stem cell proliferation and the regulation of cell development and differentiation. Very little has been discovered about the specific role for PUM1. However it has been shown to bind an mRNA sequence known as Nanos response element and to interact with RNA. PUM1 is found in heart, brain, muscle, kidney, stomach and intestine. It is not expressed in putamen, hippocampus, medulla oblangata, corpus collasum, cerebellum and caudate nucleus.

Specificity

This antibody recognizes the N-terminus of Pumilio-1 (PUM1).

Immunogen

KLH-conjugated linear peptide corresponding to the N-terminus of human Pumilio-1.

Application

RIPAb+ PUM1 Antibody detects level of RIPAb+ PUM1 & has been published & validated for use in ICC, Western Blotting, RNA Binding Protein Immunoprecipitation (RIP).

Research Sub Category
Hematopoietic Stem Cells

RNA Metabolism & Binding Proteins

Western Blot Analysis:
Representative lot data.
H9 human stem cell lysate was resolved by electrophoresis, transferred to PVDF membranes and probed with Anti-Pumilio-1 (PUM1) (1:5,000 dilution).
Proteins were visualized using a Donkey Anti-Rabbit IgG conjugated to HRP and chemiluminescence detection system.
Arrow indicates Pumilio-1 (~120 kDa).
Immunocytochemistry Analysis:
Representative lot data.
Confocal fluorescent analysis of mouse and human embryonic stem cells using Anti-Pumilio-1 (PUM1) (Red). Actin filaments have been labeled with Alexa Fluor® 488 dye - Phalloidin (Green). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytosol.

Target description

~120 kDa
UniProtKB/Swiss-Prot Q14671 (PUM1_HUMAN) entry describes 2 isoforms produced by alternative splicing: Isoform 1 at ~126 kDa and Isoform 2 at ~124 kDa

Physical form

Components:
Anti-PUM1 (Rabbit Polyclonal). One vial containing 25 µg of affinity purified polyclonal in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl,with 0.05% sodium azide before the addition of 30% glycerol. Store at -20°C.
Concentration: 0.35 mg/mL
Normal Rabbit IgG. One vial containing 125 µg Rabbit IgG in 125 µL storage buffer containing 0.05% sodium azide. Store at -20°C.
RIP Primers, p27 mRNA. One vial containing 75 μL of each primer (5 μM) specific for a region of the human p27 (Kip1/CDKN1B) mRNA. Store at -20°C.
FOR: CTG CAA CCG ACG ATT CTT CTA CT
REV: GGG CGT CTG CTC CAC AGA

Legal Information

ALEXA FLUOR is a registered trademark of Life Technologies

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

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

Packaging

10 assays per set. Recommended use: ~2.5 µg (7.2 µL) of antibody per RIP (dependent upon biological context).

Quality

RNA Binding Protein Immunoprecipitation:
Representative lot data.
RIP Lysate prepared from HeLa cells (2 X 10E7 cell equivalents per IP) were subjected to immunoprecipitation using 2.5 µg of either a normal Rabbit IgG or 2.5 µg Anti-PUM1 antibodyand the Magna RIP® RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700). Successful immunoprecipitation of PUM1-associated RNA was verified by qPCR using RIP Primers p27 mRNA, (Figure 1).
Please refer to the Magna RIP (Cat. # 17-700) or EZ-Magna RIP (Cat. # 17-701) protocol for experimental details.

Analysis Note

Control
Includes normal rabbit IgG and primers specific for human p27 mRNA.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Safety & Documentation

Safety Information

Safety Information for this product is unavailable at this time.
Protocols & Articles

Articles

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Peer-Reviewed Papers
15

References

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