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Western blot - BRIT1 antibody (ab2612)
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Western blot - BRIT1 antibody (ab2612)
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - BRIT1 antibody (ab2612)
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Product Name
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BRIT1 antibody
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See all BRIT1 antibodies (2)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to BRIT1
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Immunogen
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Synthetic peptide derived from residues 300 - 400 of Human BRIT1.
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Reacts with
(species key)
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Hu, Ms
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Specificity
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This 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.
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Tested applications
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ICC/IF, IHC-FoFr, IHC-P, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
ICC/IF: Use at a concentration of 5 µg/ml.
IHC-P: Use at an assay dependent dilution. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
IHC-FoFr: 1/300.
WB: 1/1000. Detects a band of approximately 110 kDa (predicted molecular weight: 93 kDa). Can be blocked with BRIT1 peptide (ab13737).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Cellular localization
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Centrosome.
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Research areas
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Chromatin and Nuclear Signaling >> DNA / RNA >> DNA Damage & Repair >> DNA Damage Response >> BRCT Domain Proteins
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Relevance
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BRIT1 (BRCT-repeat inhibitor of hTERT expression) contains two BRCT domains in the C terminus. It was identified in a screen for negative regulators of hTERT (human telomerase) expression. It has been implicated in chromosome condensation and DNA damage induced cellular responses. It may play a role in neurogenesis and regulation of the size of the cerebral cortex. Defects in BRIT1 are the cause of primary microcephaly 1 (MCPH1)
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Database links
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The links below go to external sites and will open in a new browser window |
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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Preservative: 0.05% Sodium Azide
Constituents: PBS, pH 7.4 Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
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Form
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Liquid
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Concentration
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0.500 mg/ml
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Storage instructions
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Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this BRIT1 antibody is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for BRIT1 antibody images, references, products related to ab2612 and other tools.
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BRIT1 antibody images:
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 Western blot - BRIT1 antibody (ab2612)
Lanes 1 - 3 : BRIT1 antibody (ab2612) at 1/250 dilution
Lanes 4 - 6 : BRIT1 antibody (ab2612) at 1/500 dilution
Lanes 7 - 9 : 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 middle band is BRIT1 (denoted by arrow) as determined by treatment with siRNA.
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Western blot - BRIT1 antibody (ab2612)
All lanes : 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
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - BRIT1 antibody (ab2612)
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 .
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Immunocytochemistry/ Immunofluorescence - BRIT1 antibody (ab2612)
ICC/IF image of ab2612 stained human Hek293 cells. The cells were PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab2612, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
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Immunohistochemistry (PFA perfusion fixed frozen sections) - BRIT1 antibody (ab2612)
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. Sophie Pezet, CNRS, Paris, France
See Abreview
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References for BRIT1 antibody (ab2612)
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This product has been used in:
Xu X et al. Microcephalin is a DNA damage response protein involved in regulation of CHK1 and BRCA1. J Biol Chem : (2004). PubMed: 15220350
If you publish research using ab2612 please let us know so that we can cite the reference on this datasheet.
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Search PubMed (MEDLINE) for references to BRIT1
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BRIT1 antibody - more information
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Customer reviews (feedback) regarding BRIT1 antibody |
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Price and availability of products related to BRIT1 antibody |
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BRIT1 antibody - related products:
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BRIT1 antibody - other tools:
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Contact Abcam with a Technical Enquiry about BRIT1 antibody |
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