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Sigma-Aldrich

RIPAb+ IGF2 mRNA-binding protein 3 - RIP Validated Antibody and Primer Set

from rabbit, purified by affinity chromatography

Synonim(y):

IGF II mRNA binding protein 3, IGF-II mRNA-binding protein 3, IGF2 mRNA-binding protein 3, KH domain containing protein overexpressed in cancer, KH domain-containing protein overexpressed in cancer, VICKZ family member 3, insulin-like growth factor 2 mRN

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About This Item

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

pochodzenie biologiczne

rabbit

Poziom jakości

klon

polyclonal

oczyszczone przez

affinity chromatography

reaktywność gatunkowa

canine, mouse, human

reaktywność gatunkowa (przewidywana na podstawie homologii)

chimpanzee (based on 100% sequence homology), dog (based on 100% sequence homology), horse (based on 100% sequence homology), rhesus monkey (based on 100% sequence homology)

producent / nazwa handlowa

RIPAb+
Upstate®

metody

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

numer dostępu NCBI

numer dostępu UniProt

Warunki transportu

dry ice

informacje o genach

human ... IGF2BP3(10643)

Opis ogólny

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.
IGF2 mRNA-binding protein 3 contains several KH domains, which are important in RNA binding and are known to be involved in RNA synthesis and metabolism.

Specyficzność

This antibody recognizes IGF2 mRNA-binding protein 3 at the C-terminus.

Immunogen

Epitope: C-terminus
KLH-conjugated linear peptide corresponding to human IGF2 mRNA-binding protein 3 at and around the C-terminus.

Zastosowanie

Immunoprecipitation from RIP lysate:
Representative lot data.
RIP lysate from HeLa cells (~2 X 10E7 cell equivalents per IP) was subjected to immunoprecipitation using 5 µg of either a normal Rabbit IgG, (Cat. # PP64B), or 5 µg of Anti-IGF2 mRNA-binding protein 3 antibody (Cat. # CS207310). Ten percent of the precipitated proteins (lane 1: Rabbit IgG, lane 2: IGF2 mRNA-binding protein 3) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-IGF2 mRNA-binding protein 3 antibody (Cat. # CS207310, 1:1000). Proteins were visualized using One-Step IP-Western kit (GenScript Cat. # L00231) Arrow indicates IGF2 mRNA-binding protein 3. The lower band in lane 2 correspond to the isoform 2.
Western Blot Analysis:
Representative lot data.
HEK293 cell lysate was resolved by electrophoresis, transferred to PVDF membranes and probed with IGF2BP3, (1:2,000) dilution. Proteins were visualized using a Donkey anti-Rabbit conjugated to HRP using a chemiluminescence detection system.
Arrow indicates IGF2BP3 (~64 kDa).
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Growth Factors & Receptors

RNA Metabolism & Binding Proteins
This RIPAb+ IGF2 mRNA-binding protein 3 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

Opakowanie

10 assays per set. Recommended use: ~5 μg of antibody per RIP (dependent upon biological context).

Jakość

RNA Binding Protein Immunoprecipitation:
Representative lot data.
RIP Lysate prepared from HepG2 cells (2 X 10E7 cell equivalents per IP) were subjected to immunoprecipitation using 5 µg of either a normal Rabbit IgG or 5 µg of Anti-IGF2 mRNA-binding protein antibody and the Magna RIP® RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of IGF2 mRNA-binding protein 3-associated RNA was verified by qPCR using RIP Primers IGF2, (Figure 1).
Please refer to the Magna RIP (Cat. # 17-700) or EZ-Magna RIP (Cat. # 17-701) protocol for experimental details.

Opis wartości docelowych

64 kDa was observed. Predicted to cross-react with all the Isoforms of IGF2BP3.

Postać fizyczna

Affinity purified
Anti-IGF2 mRNA-binding protein 3 (Rabbit Polyclonal). One vial containing 75 μL of affinity purified rabbit polyclonal antibody in PBS with 0.09% sodium azide and 30% glycerol. Store at -20°C. Concentration: 0.7 mg/mL
Normal Rabbit IgG. One vial containing 125 μg of rabbit IgG in 125 μL of storage buffer containing 0.05% sodium azide. Store at -20°C.
RIP Primer IGF2, Part # CS207311.
One vial containing 75 μL of 5 μM of each primer specific for human IGF2 mRNA. Store at -20°C.
FOR: GCG GCT TCT ACT TCA GCA G
REV: CAG GTG TCA TAT TGG AAG AAC

Przechowywanie i stabilność

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variabillity in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.

Komentarz do analizy

Control
Includes negative control normal rabbit IgG antibody and control primers specific for the cDNA of human IGF2.

Inne uwagi

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

Informacje prawne

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

Oświadczenie o zrzeczeniu się odpowiedzialności

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.
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Kod klasy składowania

12 - Non Combustible Liquids

Temperatura zapłonu (°F)

Not applicable

Temperatura zapłonu (°C)

Not applicable


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Odwiedź Bibliotekę dokumentów

Zhenzhen Wang et al.
EBioMedicine, 41, 357-369 (2019-02-25)
The cross-talk between RNA binding proteins (RBPs) and microRNAs (miRNAs) in the regulation of gene expression is a complex process. Here, we describe a new mode of regulation of TRIM25 expression mediated by an antagonistic interplay between IGF2BP3 and miR-3614-3p.

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