Overview

  • Product name

    Anti-CIRP antibody [EPR18783]
    See all CIRP primary antibodies
  • Description

    Rabbit monoclonal [EPR18783] to CIRP
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IP, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant full length protein within Human CIRP aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: Q14011

  • Positive control

    • WB: Human fetal heart and fetal kidney lysate; HeLa whole cell lysate treated at 37°C for 48 hours; HeLa whole cell lysate treated at 32°C for 48 hours; HepG2 and HCT 116 cell lysates. IHC-P: Human endometrium and breat cancer tissues. ICC/IF: HeLa and HCT 116 cells. IP: HeLa whole cell lysate.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab191885 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/1000.
IP 1/20.
WB 1/1000. Detects a band of approximately 19 kDa (predicted molecular weight: 19 kDa).
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • Function

    Cold-inducible mRNA binding protein that plays a protective role in the genotoxic stress response by stabilizing transcripts of genes involved in cell survival. Acts as a translational activator. Seems to play an essential role in cold-induced suppression of cell proliferation. Binds specifically to the 3'-untranslated regions (3'-UTRs) of stress-responsive transcripts RPA2 and TXN. Acts as a translational repressor (By similarity). Promotes assembly of stress granules (SGs), when overexpressed.
  • Tissue specificity

    Ubiquitous.
  • Sequence similarities

    Contains 1 RRM (RNA recognition motif) domain.
  • Domain

    Both the RRM domain and the arginine, glycine (RGG) rich domain are necessary for binding to the TXN 3'-untranslated region. Both the RRM domain and the arginine, glycine (RGG) rich domain (RGG repeats) are necessary for optimal recruitment into SGs upon cellular stress. The C-terminal domain containing RGG repeats is necessary for translational repression.
  • Post-translational
    modifications

    Methylated on arginine residues. Methylation of the RGG motifs is a prerequisite for recruitment into SGs.
    Phosphorylated by CK2, GSK3A and GSK3B. Phosphorylation by GSK3B increases RNA-binding activity to the TXN 3'-UTR transcript upon exposure to UV radiation.
  • Cellular localization

    Nucleus > nucleoplasm. Cytoplasm. Translocates from the nucleus to the cytoplasm after exposure to UV radiation. Translocates from the nucleus to the cytoplasm into stress granules upon various cytoplasmic stresses, such as osmotic and heat shocks. Its recruitment into stress granules occurs in the absence of TIAR proteins.
  • Information by UniProt
  • Database links

  • Alternative names

    • A18 hnRNP antibody
    • A18HNRNP antibody
    • cirbp antibody
    • CIRBP_HUMAN antibody
    • CIRP antibody
    • Cold inducible RNA binding protein antibody
    • Cold-inducible RNA-binding protein antibody
    • Glycine rich RNA binding protein antibody
    • Glycine-rich RNA-binding protein CIRP antibody
    • testicular tissue protein Li 39 antibody
    see all

Images

  • All lanes : Anti-CIRP antibody [EPR18783] (ab191885) at 1/1000 dilution

    Lane 1 : Human fetal heart lysate
    Lane 2 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 19 kDa
    Observed band size: 19 kDa


    Exposure time: 3 minutes


    5% NFDM/TBST: Blocking and diluting buffer.

  • All lanes : Anti-CIRP antibody [EPR18783] (ab191885) at 1/1000 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate treated at 37°C for 48 hours
    Lane 2 : HeLa whole cell lysate treated at 32°C for 48 hours
    Lane 3 : HCT 116 (Human colorectal carcinoma cell line) whole cell lysate
    Lane 4 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 19 kDa
    Observed band size: 19 kDa


    Exposure time: 3 minutes


    5% NFDM/TBST: Blocking and diluting buffer.

    The expression profile observed is consistent with what has been described in the literature (PMID:20735994)

  • Immunohistochemical analysis of paraffin-embedded human endometrium tissue labeling CIRP using ab191885 at 1/1000 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Counterstain: Hematoxylin.
    Inset image: negative control obtained using PBS instead of ab191885, and secondary antibody only.
    Note: Nuclear staining on normal human endometrium.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling CIRP using ab191885 at 1/1000 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Counterstain: Hematoxylin.
    Inset image: negative control obtained using PBS instead of ab191885, and secondary antibody only.
    Note: Nuclear and weak cytoplasm staining on tumor cells of breast cancer.
    Mol Carcinog. 2010 49, 130–140.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling CIRP with ab191885 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear staining on HeLa cell line.

    The nuclear counter stain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
    The negative controls are as follows:
    1. ab191885 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HCT 116 (Human colorectal carcinoma cell line) cells labeling CIRP with ab191885 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear staining on HCT 116 cell line.

    The nuclear counter stain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
    The negative controls are as follows:
    1. ab191885 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

  • Immunoprecipitation of CIRP from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate achieved using ab191885 at 1/20 dilution.
    Lane 1: Input: 10µg of HeLa whole cell lysate.
    Lane 2: HeLa whole cell lysate following IP with ab191885.
    Lane 3: negative control: IP using Rabbit monoclonal IgG (ab172730) instead of ab191885 in HeLa whole cell lysates.
    Western blot was performed using ab191885 at 1/1000 dilution.
    VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/10000 was used for detection.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST. 3 second exposure.

References

This product has been referenced in:

  • Jackson TC  et al. Infants Uniquely Express High Levels of RBM3 and Other Cold-Adaptive Neuroprotectant Proteins in the Human Brain. Dev Neurosci 40:325-336 (2018). Read more (PubMed: 30399610) »
  • Huang ZL  et al. Identification of G-Quadruplex-Binding Protein from the Exploration of RGG Motif/G-Quadruplex Interactions. J Am Chem Soc N/A:N/A (2018). Read more (PubMed: 30517002) »
See all 2 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Western blot
Sample
Bos taurus Cell lysate - whole cell (Oocytes and cumulus cells)
Gel Running Conditions
Non-reduced Denaturing (4-20%)
Loading amount
25 µg
Treatment
Cold shock (33.5 ºC) for 24h
Specification
Oocytes and cumulus cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jul 18 2019

Application
Western blot
Sample
Human Cell lysate - whole cell (HTC116 Colon Cell line)
Gel Running Conditions
Reduced Denaturing (4-12% SDS-polyacrylamide)
Loading amount
50 µg
Treatment
37ºC and 32ºC
Specification
HTC116 Colon Cell line
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C

Abcam user community

Verified customer

Submitted Feb 07 2017

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Liver)
Permeabilization
No
Specification
Liver
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Acetone

Abcam user community

Verified customer

Submitted Jun 01 2016

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