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Human Breast cancer type 2 susceptibility protein (BRCA2) ELISA Kit (HUEB1442)

SKU:
HUEB1442
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P51587
Range:
0.156-10 ng/mL
ELISA Type:
Sandwich
Synonyms:
BRCA2, Breast Cancer Susceptibility Protein 2, BRCC2
Reactivity:
Human
€649
Frequently bought together:

Description

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Human Breast cancer type 2 susceptibility protein (BRCA2) ELISA Kit

The Human Breast Cancer Type 2 Susceptibility Protein (BRCA2) ELISA Kit is specifically designed for the accurate detection of BRCA2 levels in human serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, ensuring precise and reliable results for research purposes.BRCA2 is a critical protein involved in DNA repair and maintenance, particularly in breast and ovarian cancer. Mutations in the BRCA2 gene have been linked to an increased risk of developing these types of cancer, making it a valuable biomarker for studying cancer susceptibility and potential therapeutic interventions.

This BRCA2 ELISA Kit provides researchers with a powerful tool to investigate the role of BRCA2 in cancer development and progression. With its easy-to-use format and superior performance, this kit is ideal for studies exploring the molecular mechanisms underlying breast and ovarian cancer, ultimately contributing to the advancement of personalized medicine and targeted therapies.

Product Name:Human Breast cancer type 2 susceptibility protein (BRCA2) ELISA Kit
SKU:HUEB1442
Size:96T
Target:Human Breast cancer type 2 susceptibility protein (BRCA2)
Synonyms:Fanconi anemia group D1 protein, FACD, FANCD1
Assay Type:Sandwich
Detection Method:ELISA
Reactivity:Human
Detection Range:0.156-10ng/mL
Sensitivity:0.078ng/mL
Intra CV:6.2%
Inter CV:10.9%
Linearity:
Sample1:21:41:81:16
Serum(N=5)106-116%84-96%108-118%93-102%
EDTA Plasma(N=5)100-112%108-118%87-96%99-108%
Heparin Plasma(N=5)115-115%101-111%104-114%97-109%
Recovery:
Sample TypeAverage(%)Recovery Range(%)
Serum10296-108
Plasma10498-110
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. Part of a PALB2-scaffolded HR complex containing RAD51C and which is thought to play a role in DNA repair by HR. May participate in S phase checkpoint activation. Binds selectively to ssDNA, and to ssDNA in tailed duplexes and replication fork structures. May play a role in the extension step after strand invasion at replication-dependent DNA double-strand breaks; together with PALB2 is involved in both POLH localization at collapsed replication forks and DNA polymerization activity. In concert with NPM1, regulates centrosome duplication. Interacts with the TREX-2 complex (transcription and export complex 2) subunits PCID2 and SEM1, and is required to prevent R-loop-associated DNA damage and thus transcription-associated genomic instability. Silencing of BRCA2 promotes R-loop accumulation at actively transcribed genes in replicating and non-replicating cells, suggesting that BRCA2 mediates the control of R-loop associated genomic instability, independently of its known role in homologous recombination (PubMed:24896180).
Uniprot:P51587
Sample Type:Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Specificity:Natural and recombinant human Breast cancer type 2 susceptibility protein
Sub Unit:Monomer and dimer. Interacts with RAD51; regulates RAD51 recruitment and function at sites of DNA repair. Interacts with WDR16, USP11, DMC1, ROCK2 and NPM1. Interacts with SEM1; the interaction masks a nuclear export signal in BRCA2. Interacts with both nonubiquitinated and monoubiquitinated FANCD2; this complex also includes XRCC3 and phosphorylated FANCG. Part of a BRCA complex containing BRCA1, BRCA2 and PALB2. Interacts directly with PALB2 which may serve as a scaffold for a HR complex containing PALB2, BRCA2, RAD51C, RAD51 and XRCC3. Interacts with BRCA1 only in the presence of PALB2 which serves as the bridging protein. Interacts with POLH; the interaction is direct. Interacts with the TREX-2 complex subunits PCID2 and SEM1 (PubMed:24896180, PubMed:21719596).
Research Area:Cancer
Subcellular Location:Nucleus Cytoplasm Cytoskeleton Microtubule organizing center Centrosome
Storage:Please see kit components below for exact storage details
Note:For research use only
UniProt Protein Function:BRCA2: involved in double-strand break repair and/or homologous recombination. May participate in S phase checkpoint activation. Interacts with RAD51 and DSS1. Interacts with ubiquitinated FANCD2. Interacts with PALB2, enables the recombinational repair and checkpoint functions. Interacts with WDR16. Defects in BRCA2 are a cause of genetic susceptibility to breast cancer and may underlie susceptibility to uveal melanoma.
UniProt Protein Details:

Protein type:Tumor suppressor; Nuclear receptor co-regulator; DNA repair, damage

Chromosomal Location of Human Ortholog: 13q12.3

Cellular Component: nucleoplasm; centrosome; protein complex; cytoplasm; BRCA2-MAGE-D1 complex; nucleus; secretory granule

Molecular Function:gamma-tubulin binding; protein binding; H4 histone acetyltransferase activity; histone acetyltransferase activity; protease binding; H3 histone acetyltransferase activity; single-stranded DNA binding

Biological Process: positive regulation of transcription, DNA-dependent; cytokinesis; cell aging; positive regulation of mitotic cell cycle; DNA repair; regulation of cytokinesis; oocyte maturation; inner cell mass cell proliferation; negative regulation of mammary gland epithelial cell proliferation; response to UV-C; double-strand break repair via homologous recombination; DNA damage response, signal transduction by p53 class mediator resulting in transcription of p21 class mediator; male meiosis I; nucleotide-excision repair; double-strand break repair; response to gamma radiation; hemopoiesis; spermatogenesis; replication fork protection; brain development; DNA damage response, signal transduction by p53 class mediator resulting in induction of apoptosis; centrosome duplication; female gonad development; response to X-ray

Disease: Pancreatic Cancer, Susceptibility To, 2; Breast-ovarian Cancer, Familial, Susceptibility To, 2; Prostate Cancer; Breast Cancer; Medulloblastoma; Glioma Susceptibility 3; Tracheoesophageal Fistula With Or Without Esophageal Atresia; Fanconi Anemia, Complementation Group D1; Wilms Tumor 1

NCBI Summary:Inherited mutations in BRCA1 and this gene, BRCA2, confer increased lifetime risk of developing breast or ovarian cancer. Both BRCA1 and BRCA2 are involved in maintenance of genome stability, specifically the homologous recombination pathway for double-strand DNA repair. The BRCA2 protein contains several copies of a 70 aa motif called the BRC motif, and these motifs mediate binding to the RAD51 recombinase which functions in DNA repair. BRCA2 is considered a tumor suppressor gene, as tumors with BRCA2 mutations generally exhibit loss of heterozygosity (LOH) of the wild-type allele. [provided by RefSeq, Dec 2008]
UniProt Code:P51587
NCBI GenInfo Identifier:14424438
NCBI Gene ID:675
NCBI Accession:P51587.2
UniProt Secondary Accession:P51587,O00183, O15008, Q13879, Q5TBJ7,
UniProt Related Accession:P51587
Molecular Weight:3418
NCBI Full Name:Breast cancer type 2 susceptibility protein
NCBI Synonym Full Names:breast cancer 2, early onset
NCBI Official Symbol:BRCA2
NCBI Official Synonym Symbols:FAD; FACD; FAD1; GLM3; BRCC2; FANCD; PNCA2; FANCD1; XRCC11; BROVCA2
NCBI Protein Information:breast cancer type 2 susceptibility protein; Fanconi anemia group D1 protein; breast cancer 2 tumor suppressor; BRCA1/BRCA2-containing complex, subunit 2; breast and ovarian cancer susceptibility gene, early onset
UniProt Protein Name:Breast cancer type 2 susceptibility protein
UniProt Synonym Protein Names:Fanconi anemia group D1 protein
Protein Family:BRCA2 and CDKN1A-interacting protein
UniProt Gene Name:BRCA2
UniProt Entry Name:BRCA2_HUMAN
Component Quantity (96 Assays) Storage
ELISA Microplate (Dismountable) 8×12 strips -20°C
Lyophilized Standard 2 -20°C
Sample Diluent 20ml -20°C
Assay Diluent A 10mL -20°C
Assay Diluent B 10mL -20°C
Detection Reagent A 120µL -20°C
Detection Reagent B 120µL -20°C
Wash Buffer 30mL 4°C
Substrate 10mL 4°C
Stop Solution 10mL 4°C
Plate Sealer 5 -

Other materials and equipment required:

  • Microplate reader with 450 nm wavelength filter
  • Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
  • Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Buffer resevoir

*Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Allow all reagents to reach room temperature (Please do not dissolve the reagents at 37°C directly). All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Keep appropriate numbers of strips for 1 experiment and remove extra strips from microtiter plate. Removed strips should be resealed and stored at -20°C until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their experiments. We recommend running all samples in duplicate.

Step
1. Add Sample: Add 100µL of Standard, Blank, or Sample per well. The blank well is added with Sample diluent. Solutions are added to the bottom of micro ELISA plate well, avoid inside wall touching and foaming as possible. Mix it gently. Cover the plate with sealer we provided. Incubate for 120 minutes at 37°C.
2. Remove the liquid from each well, don't wash. Add 100µL of Detection Reagent A working solution to each well. Cover with the Plate sealer. Gently tap the plate to ensure thorough mixing. Incubate for 1 hour at 37°C. Note: if Detection Reagent A appears cloudy warm to room temperature until solution is uniform.
3. Aspirate each well and wash, repeating the process three times. Wash by filling each well with Wash Buffer (approximately 400µL) (a squirt bottle, multi-channel pipette,manifold dispenser or automated washer are needed). Complete removal of liquid at each step is essential. After the last wash, completely remove remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper.
4. Add 100µL of Detection Reagent B working solution to each well. Cover with the Plate sealer. Incubate for 60 minutes at 37°C.
5. Repeat the wash process for five times as conducted in step 3.
6. Add 90µL of Substrate Solution to each well. Cover with a new Plate sealer and incubate for 10-20 minutes at 37°C. Protect the plate from light. The reaction time can be shortened or extended according to the actual color change, but this should not exceed more than 30 minutes. When apparent gradient appears in standard wells, user should terminatethe reaction.
7. Add 50µL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing.
8. Determine the optical density (OD value) of each well at once, using a micro-plate reader set to 450 nm. User should open the micro-plate reader in advance, preheat the instrument, and set the testing parameters.
9. After experiment, store all reagents according to the specified storage temperature respectively until their expiry.

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample Type Protocol
Serum If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.
Plasma Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.
Urine & Cerebrospinal Fluid Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
Cell culture supernatant Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
Cell lysates Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Tissue homogenates The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.
Tissue lysates Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Breast Milk Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.
ELISA
Human Breast cancer type 2 susceptibility protein / BRCA2 ELISA Kit
Human BRCA2 (Breast Cancer Susceptibility Protein 2) CLIA Kit (HUES00402)

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