Overview

  • Product nameAnti-BRIT1 antibody
    See all BRIT1 primary antibodies
  • Description
    Rabbit polyclonal to BRIT1
  • SpecificityThis antibody detects a band of ~100kD in U2OS and HEK293 cells (Xu et al. 2004). The band is reduced in U2OS cells treated with BRIT1 siRNA (Xu et al. 2004) showing that the antibody is specific for BRIT1. Some batches of this antibody may also recognise additional isoforms.
  • Tested applicationsSuitable for: IHC-P, IHC-FoFr, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide corresponding to Human BRIT1 aa 400-500.
    (Peptide available as ab13737, ab13737, ab13737, ab13737)

  • Positive control
    • This antibody gave a positive signal in Hela, Jurkat and HEK293 whole cell lysates and Hela whole cell lysate Bleomycin Treated (40U/ml) and Hydroxyurea Treated (48hr, 1uM)

Properties

Applications

Our Abpromise guarantee covers the use of ab2612 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
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
IHC-FoFr 1/300.
WB 1/500 - 1/1000. Detects a band of approximately 110 kDa (predicted molecular weight: 93 kDa).

Target

  • FunctionImplicated in chromosome condensation and DNA damage induced cellular responses. May play a role in neurogenesis and regulation of the size of the cerebral cortex.
  • Tissue specificityExpressed in fetal brain, liver and kidney.
  • Involvement in diseaseDefects in MCPH1 are the cause of microcephaly primary type 1 (MCPH1) [MIM:251200]; also known as true microcephaly or microcephaly vera. Microcephaly is defined as a head circumference more than 3 standard deviations below the age-related mean. Brain weight is markedly reduced and the cerebral cortex is disproportionately small. Despite this marked reduction in size, the gyral pattern is relatively well preserved, with no major abnormality in cortical architecture. Primary microcephaly is further defined by the absence of other syndromic features or significant neurological deficits. This entity is inherited as autosomal recessive trait.
  • Sequence similaritiesContains 3 BRCT domains.
  • Cellular localizationCytoplasm > cytoskeleton > centrosome.
  • Information by UniProt
  • Database links
  • Alternative names
    • BRCT repeat inhibitor of TERT expression 1 antibody
    • BRIT 1 antibody
    • FLJ12847 antibody
    • Hypothetical protein FLJ12847 antibody
    • MCPH 1 antibody
    • MCPH1 antibody
    • MCPH1_HUMAN antibody
    • MCT antibody
    • Microcephalin 1 antibody
    • Microcephalin antibody
    • Microcephaly primary autosomal recessive 1 antibody
    see all

Anti-BRIT1 antibody images

  • Lanes 1 - 3 : Anti-BRIT1 antibody (ab2612) at 1/250 dilution
    Lanes 4 - 6 : Anti-BRIT1 antibody (ab2612) at 1/500 dilution
    Lanes 7 - 9 : Anti-BRIT1 antibody (ab2612) at 1/1000 dilution

    Lane 1 : 30 ug of LCL1 whole cell lysate.
    Lane 2 : 30 ug of LCL2 whole cell lysate.
    Lane 3 : 30 ug of LCL3 whole cell lysate.
    Lane 4 : 30 ug of LCL1 whole cell lysate.
    Lane 5 : 30 ug of LCL2 whole cell lysate.
    Lane 6 : 30 ug of LCL3 whole cell lysate.
    Lane 7 : 30 ug of LCL1 whole cell lysate.
    Lane 8 : 30 ug of LCL2 whole cell lysate.
    Lane 9 : 30 ug of LCL3 whole cell lysate.


    Performed under reducing conditions.

    Predicted band size : 93 kDa
    Observed band size : 95 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 180 kDa,60 kDa (possible cleavage fragment),70 kDa (possible cleavage fragment). We are unsure as to the identity of these extra bands.

    Exposure time : 15 seconds

    LCL1, LCL2 and LCL3 are three patient derived fibroblast cell lines.

    The indicated dilution of antibody was incubated with 30 µg of whole cell extract.

    The middle band is BRIT1 (this has been confirmed by siRNA analysis).

    15 second exposure.

    LCL1, LCL2, and LCL3 are three patient derived fibroblast cell lines. The middle band is BRIT1 (denoted by arrow) as determined by treatment with siRNA.
  • All lanes : Anti-BRIT1 antibody (ab2612) at 1/1000 dilution

    Lane 1 : GUS (beta glucuronidase) expression negative control.
    Lane 2 : In vitro expressed human BRIT1

    Secondary
    A rabbit polyclonal to Goat IgG H&L (HRP) secondary antibody was used at a 1:5000 dilution.
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 93 kDa
    Observed band size : 100 kDa (why is the actual band size different from the predicted?)
    ab2612 was incubated for one hour at room temperature with the membrane in 5% non-fat dry milk plus PBST.

    This image was submitted courtesy of I.Gavvovidis, D. Schindler, Institut fuer Humangenetik, Universitaet Wuerzburg
  • Image courtesy of Human Protein Atlas

    ab2612 staining in human lung, showing staining of the surface epithelial cells (in brown). Paraffin embedded lung tissue was incubated with ab2612 (1:100 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab2612 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org

    .

  • Immunohistochemistical detection of BRIT1 using antibody (ab2612) on paraformaldehyde perfusion fixed mouse heart tissue sections. Primary antibody diluted @ 1/300 & incubated for 18 hours @ 20°C in PBS + 0.3 % Triton X100. Secondary antibody: goat anti-rabbit conjugated to biotin (1/300).  The antibody produced a weak cytoplasmic staining in cardiac cells.

    See Abreview

  • All lanes : Anti-BRIT1 antibody (ab2612) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Human BRIT1 peptide (ab13737) at 1 µg/ml
    Lane 5 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate with Human BRIT1 peptide (ab13737) at 1 µg/ml
    Lane 6 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate with Human BRIT1 peptide (ab13737) at 1 µg/ml

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 93 kDa
    Observed band size : 105 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 55-60 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 3 minutes
  • All lanes : Anti-BRIT1 antibody (ab2612) at 1 µg/ml (BLOCKED IN 5% MILK)

    Lane 1 : Hela Whole Cell Lysate - Bleomycin Treated (40U/ml)
    Lane 2 : Hela Whole Cell Lysate - Hydroxyurea Treated (48hr, 1uM)

    Lysates/proteins at 25 µg per lane.

    Secondary
    Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size : 93 kDa
    Observed band size : 105 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 19 kDa,55 kDa,60 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 12 minutes

References for Anti-BRIT1 antibody (ab2612)

This product has been referenced in:
  • Ge C  et al. The UBC Domain Is Required for BRUCE to Promote BRIT1/MCPH1 Function in DSB Signaling and Repair Post Formation of BRUCE-USP8-BRIT1 Complex. PLoS One 10:e0144957 (2015). Read more (PubMed: 26683461) »
  • Alsiary R  et al. Deregulation of microcephalin and ASPM expression are correlated with epithelial ovarian cancer progression. PLoS One 9:e97059 (2014). IHC-P ; Human . Read more (PubMed: 24830737) »

See all 8 Publications for this product

Product Wall

Thank you for your patience.

I can now confirm that ab82629 was tested with partly purified SPNS2 from liver tissue.

We recommend as positive control for endogenous SPNS2 also embryonic cells or mature liver cells (FaO, HepG2).
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I would like to reassure you that both antibodiesare tested and covered by our guarantee for WB.

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Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (Brain)
Specification Brain
Fixative Paraformaldehyde
Permeabilization Yes - Triton-X
Blocking step 0.1 M glycine buffer as blocking agent for 30 minute(s) · Concentration: 7.5µg/mL · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted May 25 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Heart)
Specification Heart
Fixative Paraformaldehyde
Antigen retrieval step None
Permeabilization No
Username

Dr. Sophie Pezet

Verified customer

Submitted Mar 17 2010

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