Recombinant
RabMAb

Anti-IRF1 antibody [EPR18301] (ab186384)

Overview

  • Product name
    Anti-IRF1 antibody [EPR18301]
    See all IRF1 primary antibodies
  • Description
    Rabbit monoclonal [EPR18301] to IRF1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, WB, Flow Cyt, IP, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human IRF1 aa 100 to the C-terminus. The exact sequence is proprietary.
    Database link: P10914

  • Positive control
    • WB: Jurkat and IFN-gamma treated HeLa whole cell lysate; Mouse brain, heart and spleen lysates; rat brain and heart lysates; MOLT-4, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates. IHC-P: Human colon and Human gastric adenocarcinoma tissue. ICC/IF: MOLT-4, Jurkat, NIH\3T3, C6 cells. Flow Cytometry: Jurkat cells. IP: Jurkat 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

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EPR18301
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab186384 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 1/500.
WB 1/1000. Detects a band of approximately 48 kDa (predicted molecular weight: 37 kDa).
Flow Cyt 1/250.
IP 1/80.
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • Function
    Specifically binds to the upstream regulatory region of type I IFN and IFN-inducible MHC class I genes (the interferon consensus sequence (ICS)) and activates those genes. Acts as a tumor suppressor.
  • Involvement in disease
    Defects in IRF1 are a cause of gastric cancer (GASC) [MIM:613659]; also called gastric cancer intestinal or stomach cancer. Gastric cancer is a malignant disease which starts in the stomach, can spread to the esophagus or the small intestine, and can extend through the stomach wall to nearby lymph nodes and organs. It also can metastasize to other parts of the body. The term gastric cancer or gastric carcinoma refers to adenocarcinoma of the stomach that accounts for most of all gastric malignant tumors. Two main histologic types are recognized, diffuse type and intestinal type carcinomas. Diffuse tumors are poorly differentiated infiltrating lesions, resulting in thickening of the stomach. In contrast, intestinal tumors are usually exophytic, often ulcerating, and associated with intestinal metaplasia of the stomach, most often observed in sporadic disease.
  • Sequence similarities
    Belongs to the IRF family.
    Contains 1 IRF tryptophan pentad repeat DNA-binding domain.
  • Post-translational
    modifications
    Sumoylation represses the transcriptional activity and displays enhanced resistance to protein degradation. Inactivates the tumor suppressor activity. Elevated levels in tumor cells. Major site is Lys-275. Sumoylation is enhanced by PIAS3 (By similarity). Desumoylated by SENP1 in tumor cells and appears to compete with ubiquitination on C-terminal sites.
    Ubiquitinated. Appears to compete with sumoylation on C-terminal sites.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • Interferon regulatory factor 1 antibody
    • Interferon regulatory factor 1 isoform +I9 antibody
    • Interferon regulatory factor 1 isoform d78 antibody
    • Interferon regulatory factor 1 isoform delta4 antibody
    • Interferon regulatory factor 1 isoform delta7 antibody
    • IRF 1 antibody
    • IRF-1 antibody
    • IRF1 antibody
    • IRF1_HUMAN antibody
    • MAR antibody
    • MAR1 antibody
    see all

Images

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MOLT-4 (Human lymphoblastic leukemia) cells labeling IRF1 with ab186384 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

    Confocal image showing nuclear and cytoplasmic staining on MOLT-4 cells.

    The nuclear counterstain 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 1/500 dilution (red).
    The negative controls are as follows:-
    -ve control 1 - ab186384 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia from peripheral blood) cells labeling IRF1 with ab186384 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

    Confocal image showing cytoplasmic and weakly nuclear staining on Jurkat cells.

    The nuclear counterstain 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 1/500 dilution (red).
    The negative controls are as follows:-
    -ve control 1 - ab186384 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embyro fibroblast cells) cells labeling IRF1 with ab186384 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

    Confocal image showing cytoplasmic and weakly nuclear staining on NIH/3T3 cells.

    The nuclear counterstain 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 1/500 dilution (red).
    The negative controls are as follows:-
    -ve control 1 - ab186384 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

     

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C6 (Rat glial tumor cells) cells labeling IRF1 with ab186384 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

    Confocal image showing cytoplasmic and nuclear staining on C6 cells.

    The nuclear counterstain 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 1/500 dilution (red).
    The negative controls are as follows:-
    -ve control 1 - ab186384 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

     

  • All lanes : Anti-IRF1 antibody [EPR18301] (ab186384) at 1/10000 dilution

    Lane 1 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
    Lane 2 : Untreated HeLa (Human epithelial cells from cervix adenocarcinoma ) whole cell lysate
    Lane 3 : HeLa whole cell lysates treated with 10 ng/ml IFN-gamma for 24 hours

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 37 kDa
    Observed band size: 48 kDa (why is the actual band size different from the predicted?)


    Exposure time: 15 seconds


    Blocking/dilution buffer: 5% NFDM/TBST.

  • Anti-IRF1 antibody [EPR18301] (ab186384) at 1/1000 dilution + MOLT-4 (Human lymphoblastic leukemia) whole cell lysate 20ug

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 37 kDa
    Observed band size: 48 kDa (why is the actual band size different from the predicted?)


    Exposure time: 30 seconds


    Blocking/dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-IRF1 antibody [EPR18301] (ab186384) at 1/1000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Mouse spleen lysate
    Lane 4 : Rat brain lysate
    Lane 5 : Rat heart lysate
    Lane 6 : C6 (rat glioma tumor) whole cell lysate
    Lane 7 : RAW 264.7 (mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 8 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 9 : NIH/3T3 (mouse embryo fibroblast) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 37 kDa
    Observed band size: 48 kDa (why is the actual band size different from the predicted?)


    Exposure time: 30 seconds


    Blocking/dilution buffer: 5% NFDM/TBST.

  • Flow cytometric analysis of 2% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells fromperipheral blood) cells labeling IRF1 with ab186384 at 1/250 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

  • Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling IRF1 with ab186384 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nuclear and cytoplasmic staining on epithelial cells of normal Human colon is observed.

    Counter stained with Hematoxylin.
    Negative control: Used PBS instead of primary ab, secondary ab is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).

  • Immunohistochemical analysis of paraffin-embedded Human gastric adenocarcinoma tissue labeling IRF1 with ab186384 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nuclear and weak cytoplasmic staining on tumor cells of gastric adenocarcinoma is observed.

    Counter stained with Hematoxylin.
    Negative control: Used PBS instead of primary ab, secondary ab is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).

  • IRF1 was immunoprecipitated from 1mg of Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate with ab186384 at 1/80 dilution.

    Lane 1: Input Jurkat whole cell extract (10µg). 

    Lane 2: Jurkat whole cell lysate following immunoprecipitation with ab186384.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab186384 in Jurkat whole cell lysate.

    Western blot was performed from the immunoprecipitate using ab186384 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used as secondary antibody at 1/1500 dilution.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST. 30 second exposure.

References

This product has been referenced in:
  • Chen X  et al. Specific microRNA signatures responsible for immune disturbance related to hip fracture in aged rats. J Orthop Surg Res 13:17 (2018). WB ; Rat . Read more (PubMed: 29357879) »
  • Campos Carrascosa L  et al. Reciprocal regulation of the Il9 locus by counteracting activities of transcription factors IRF1 and IRF4. Nat Commun 8:15366 (2017). CHIPseq ; Mouse . Read more (PubMed: 28497800) »

See all 2 Publications for this product

Customer reviews and Q&As

Abcam has not validated the combination of species/application used in this Abreview.
Application
ChIP
Sample
Human Cell lysate - nuclear (PH5CH8)
Negative control
IgG control ChIP
Specification
PH5CH8
Detection step
Other
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 5 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde
Username

Abcam user community

Verified customer

Submitted Dec 07 2017

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Primary mouse neurons)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
30 µg
Treatment
IFN-gamma for 48 hrs
Specification
Primary mouse neurons
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Sep 20 2017

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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