Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab9143 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 Use at an assay dependent concentration. PubMed: 12915460
IP Use a concentration of 1 - 4 µg/ml.
WB 1/1000 - 1/10000. Detects a band of approximately 380 kDa (predicted molecular weight: 380 kDa).

Target

  • Function
    Involved in double-strand break repair and/or homologous recombination. Binds RAD51 and potentiates recombinational DNA repair by promoting assembly of RAD51 onto single-stranded DNA (ssDNA). Acts by targeting RAD51 to ssDNA over double-stranded DNA, enabling RAD51 to displace replication protein-A (RPA) from ssDNA and stabilizing RAD51-ssDNA filaments by blocking ATP hydrolysis. May participate in S phase checkpoint activation. Binds selectively to ssDNA, and to ssDNA in tailed duplexes and replication fork structures.
  • Tissue specificity
    Highest levels of expression in breast and thymus, with slightly lower levels in lung, ovary and spleen.
  • Involvement in disease
    Defects in BRCA2 are a cause of susceptibility to breast cancer (BC) [MIM:114480]. A common malignancy originating from breast epithelial tissue. Breast neoplasms can be distinguished by their histologic pattern. Invasive ductal carcinoma is by far the most common type. Breast cancer is etiologically and genetically heterogeneous. Important genetic factors have been indicated by familial occurrence and bilateral involvement. Mutations at more than one locus can be involved in different families or even in the same case.
    Defects in BRCA2 are the cause of pancreatic cancer type 2 (PNCA2) [MIM:613347]. It is a malignant neoplasm of the pancreas. Tumors can arise from both the exocrine and endocrine portions of the pancreas, but 95% of them develop from the exocrine portion, including the ductal epithelium, acinar cells, connective tissue, and lymphatic tissue.
    Defects in BRCA2 are a cause of susceptibility to breast-ovarian cancer familial type 2 (BROVCA2) [MIM:612555]. A condition associated with familial predisposition to cancer of the breast and ovaries. Characteristic features in affected families are an early age of onset of breast cancer (often before age 50), increased chance of bilateral cancers (cancer that develop in both breasts, or both ovaries, independently), frequent occurrence of breast cancer among men, increased incidence of tumors of other specific organs, such as the prostate.
    Defects in BRCA2 are the cause of Fanconi anemia complementation group D type 1 (FANCD1) [MIM:605724]. It is a disorder affecting all bone marrow elements and resulting in anemia, leukopenia and thrombopenia. It is associated with cardiac, renal and limb malformations, dermal pigmentary changes, and a predisposition to the development of malignancies. At the cellular level it is associated with hypersensitivity to DNA-damaging agents, chromosomal instability (increased chromosome breakage) and defective DNA repair.
    Defects in BRCA2 are a cause of glioma type 3 (GLM3) [MIM:613029]. Gliomas are benign or malignant central nervous system neoplasms derived from glial cells. They comprise astrocytomas and glioblastoma multiforme that are derived from astrocytes, oligodendrogliomas derived from oligodendrocytes and ependymomas derived from ependymocytes.
  • Sequence similarities
    Contains 8 BRCA2 repeats.
  • Post-translational
    modifications
    Phosphorylated by ATM upon irradiation-induced DNA damage.
    Ubiquitinated in the absence of DNA damage; this does not lead to proteasomal degradation. In contrast, ubiquitination in response to DNA damage leads to proteasomal degradation.
  • Information by UniProt
  • Database links
  • Alternative names
    • BRCA 2 antibody
    • BRCA1/BRCA2 containing complex subunit 2 antibody
    • Brca2 antibody
    • BRCA2, DNA repair associated antibody
    • BRCA2_HUMAN antibody
    • BRCC 2 antibody
    • BRCC2 antibody
    • Breast and ovarian cancer susceptibility gene early onset antibody
    • breast and ovarian cancer susceptibility protein 2 antibody
    • Breast cancer 2 early onset antibody
    • Breast Cancer 2 tumor suppressor antibody
    • Breast cancer susceptibility protein BRCA2 antibody
    • Breast cancer type 2 susceptibility protein antibody
    • BROVCA2 antibody
    • FACD antibody
    • FAD 1 antibody
    • FAD antibody
    • FAD1 antibody
    • FANCB antibody
    • FANCD 1 antibody
    • FANCD antibody
    • FANCD1 antibody
    • FANCD1 gene antibody
    • Fanconi anemia complementation group D1 antibody
    • Fanconi anemia group D1 protein antibody
    • GLM3 antibody
    • mutant BRCA2 antibody
    • OTTHUMP00000018803 antibody
    • OTTHUMP00000042401 antibody
    • PNCA2 antibody
    • XRCC11 antibody
    see all

Images

  • ab9143 immunoprecipitating BRCA2 at 6 µg per reaction.

    Lane 1: ab9143 in HEK2936 cell lysate.

    Lane 2: Rabbit polyclonal antibody (ab123491) used instead of ab9143 in 293T cell lysate

    Lane 3 (control): Secondary antibody only control

    For blotting, ab9143 immunoprecipitated BRAC2 at 1 µg/ml. Detected by Chemiluminescence.

References

This product has been referenced in:
  • Orthwein A  et al. A mechanism for the suppression of homologous recombination in G1 cells. Nature 528:422-6 (2015). Read more (PubMed: 26649820) »
  • Suzuki S  et al. Inhibition of post-translational N-glycosylation by HRD1 that controls the fate of ABCG5/8 transporter. Sci Rep 4:4258 (2014). Human . Read more (PubMed: 24584735) »
See all 3 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (293T)
Gel Running Conditions
Reduced Denaturing (4-20%)
Loading amount
50 µg
Specification
293T

Abcam user community

Verified customer

Submitted Nov 02 2016

Answer

Thanks for your inquiry. I have copied below some details on transferring large proteins. Detailed instructions for the transfer process can be found on the websites of the manufacturers of transfer apparatus, and will vary depending on the system. The principle is the same in each case, though. Just as proteins with an electrical charge (provided by the SDS bound to them) can be induced to travel through a gel in an electrical field, so can the proteins be transferred in an electrical field from the gel onto a sturdy support, a membrane that “blots” the proteins from the gel. (Early methods relied on diffusion; blotting in an electrical field is now standard). Transfer can be done in wet or semi-dry conditions. Semi-dry transfer is generally faster but wet transfer is a less prone to failure due to drying of the membrane and is especially recommended for large proteins, >100 kD. For both kinds of transfer, the membrane is placed next to the gel. The two are sandwiched between absorbent materials, and the sandwich is clamped between solid supports to maintain tight contact between the gel and membrane. In wet transfer, the gel and membrane are sandwiched between sponge and paper (sponge/paper/gel/membrane/paper/sponge) and all are clamped tightly together after ensuring no air bubbles have formed between the gel and membrane. The sandwich is submerged in transfer buffer to which an electrical field is applied. The negatively-charged proteins travel towards the positively-charged electrode, but the membrane stops them, binds them, and prevents them from continuing on. A standard buffer for wet transfer is the same as the 1X Tris-glycine buffer used for the migration/running buffer without SDS but with the addition of methanol to a final concentration of 20%. For proteins larger than 80 kD, it is recommended that SDS is included at a final concentration of 0.1%. Two types of membranes are available: nitrocellulose and PVDF (positively-charged nylon). The choice is personal and both work very well. PVDF membranes require careful pre-treatment: cut the membrane to the appropriate size then soak it in methanol for 1-2 min. Incubate in ice cold transfer buffer for 5 minutes. The gel needs to equilibrate for 3-5 minutes in ice cold transfer buffer. Failure to do so will cause shrinking while transferring, and a distorted pattern of transfer. Note on transfer of large and small proteins The balance of SDS and methanol in the transfer buffer, protein size, and gel percentage can affect transfer efficiency. The following modifications will encourage efficient transfer. Large proteins (>100 kD) For large proteins, transfer out of the gel may be very slow, just as they run slowly within the gel during separation. If blotting a large protein, be sure to run your samples in a low-concentration gel, 8% or less. These will be very fragile, so handle carefully. Large proteins will tend to precipitate in the gel, hindering transfer. Adding SDS to a final concentration of 0.1% in the transfer buffer will discourage this. Methanol tends to remove SDS from proteins, so reducing the methanol percentage to 10% or less will also guard against precipitation. Lowering methanol in the transfer buffer also promotes swelling of the gel, allowing large proteins to transfer more easily. Methanol is only necessary if using nitrocellulose. If using PVDF, methanol can be removed from the transfer buffer altogether, and is only needed to activate the PVDF before assembling the gel/membrane sandwich. Choose wet transfer overnight at 4°C instead of semi-dry transfer. I hope you find this information useful. Please feel free to contact us if you require further assistance.

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Answer

Thank you for your enquiry and your interest in our products. We would like to confirm that the two product are the same and from the same originator. The only difference is that ab1292 is Rabbit polyclonal to BRCA2 (Agarose) suitable for only IP application.

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

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