BRCA1 ELISA Kit

Catalog No. ABIN8068089

This Colorimetric ELISA kit is designed for the quantitative measurement of Human BRCA1.

Quick Overview for BRCA1 ELISA Kit (ABIN8068089)

Target: BRCA1 (Breast Cancer 1 (BRCA1))

Binding Specificity: total, ubiquitinated

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Sample Type: Cell Lysate, Tissue Lysate

Product Details BRCA1 ELISA Kit

Purpose: Measures ubiquitinated BRCA1 and total BRCA1 in human cell and tissue lysates, enabling precise assessment of BRCA1 ubiquitination by ELISA. This assay can be developed for other species (mouse, rat, etc.) upon request. Contact techsupport@raybiotech.com.

Analytical Method: Quantitative

Components:

Size / Qty
• Microplate Coated With Anti-Pan-BRCA1 Antibody
• Anti-Ubiquitination Detection Antibody
• Anti-BRCA1 Detection Antibody
• HRP-Conjugated Streptavidin
• 200 μL
• Wash Buffer
• Assay Diluent
• Lysis Buffer
• TMB One-Step Substrate Reagent
• Stop Solution

Material not included:

• Microplate reader capable of measuring absorbance at 450 nm
• Protease and Phosphatase inhibitors
• Precision pipettes to deliver 2 μL to 1 mL volumes
• Adjustable 1-25 mL pipettes for reagent preparation
• 100 mL and 1 liter graduated cylinders
• Log-log graph paper or computer and software for ELISA data analysis
• Absorbent paper
• Tubes to prepare sample dilutions
• Distilled or deionized water
• Shaker

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Plate: Pre-coated

Protocol:

1. Prepare all reagents and samples as instructed.
2. Add 100 μL sample to each well. Incubate 2.5 hours at room temperature or overnight at 4 °C with gentle shaking.
3. Add 100 μL prepared detection antibody to each well. Incubate for 1 hour at room temperature with gentle shaking.
4. Add 100 μL prepared HRP-Conjugated Streptavidin solution. Incubate for 1 hour at room temperature with gentle shaking.
5. Add 100 μL TMB One-Step Substrate Reagent to each well. Incubate 30 minutes at room temperature.
6. Add 50 μL Stop Solution to each well. Read at 450 nm immediately.

Restrictions: For Research Use only

Handling

Storage: -20 °C

Storage Comment: The entire kit may be stored at -20°C for up to 6 months from the date of shipment. Avoid repeated freeze-thaw cycles.

Target Details for BRCA1

Target: BRCA1 (Breast Cancer 1 (BRCA1))

Alternative Name: BRCA1

Background:

Gene Symbols: BRCA1, RNF53

Background: The Ubiquitinated-BRCA1 and Total-BRCA1 ELISA Kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human ubiquitinated ATP-citrate synthase and total ATP-citrate synthase. An anti-pan BRCA1 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and BRCA1 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-ubiquitination antibody is used to detect ubiquitinated BRCA1 or biotinylated anti-BRCA1 antibody is used to detect total BRCA1 . After washing away unbound antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of ubiquitinated BRCA1 or total BRCA1 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

Protein Name (Synonyms): Breast cancer type 1 susceptibility protein, RING finger protein 53, RING-type E3 ubiquitin transferase BRCA1

Gene ID: 672

UniProt: P38398

Pathways: Cell Division Cycle, DNA Damage Repair, Intracellular Steroid Hormone Receptor Signaling Pathway, Positive Regulation of Response to DNA Damage Stimulus