BRAF ELISA Kit

Catalog No. ABIN5652152

Human BRAF ELISA Kit, Colorimetric assay for quantification of Human BRAF.

Quick Overview for BRAF ELISA Kit (ABIN5652152)

Target: BRAF (B-Raf proto-oncogene, serine/threonine kinase (BRAF))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0.312 ng/mL - 20 ng/mL

Application: ELISA

Sample Type: Cell Lysate, Tissue Homogenate

Product Details BRAF ELISA Kit

Minimum Detection Limit: 0.312 ng/mL

Purpose: Human B-Raf Proto Oncogene Serine/Threonine Protein Kinase (BRAF) ELISA Kit for detecting BRAF in tissue homogenates, cell lysates

Analytical Method: Quantitative

Specificity: This assay has high sensitivity and excellent specificity for detection of B-Raf Proto Oncogene Serine/Threonine Protein Kinase (BRAF). No significant cross-reactivity or interference between B-Raf Proto Oncogene Serine/Threonine Protein Kinase (BRAF) and analogues was observed.

Sensitivity: 0.115 ng/mL

Components:

• Pre-coated, ready to use 96-well strip plate
• Standar
• Detection Reagent A
• Detection Reagent B
• TMB Substrate
• Stop Solution
• Assay Diluent A
• Assay Diluent B
• Wash Buffer (30x concentrate)
• Standard Diluent
• Plate Sealer
• Instruction Manual

Material not included:

• Microplate Reader with 450 nm filter
• Single or multi-channel pipettes with high precision and disposable tips
• Microcentrifuge tubes
• Deionized or distilled watter
• Absorbent paper for blotting the microplate
• Container for wash solution
• 0.01 M (or 1x) Phosphate Buffered Saline (PBS), ph 7.0-7.2

Application Details

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

Comment:

This product is tested and assembled before shipping to ensure that it meets its quality specifications. All kits are tested to confirm that they fall within their defined Inter- and Intra- assay coefficient of variation.

Sample Volume: 100 μL

Assay Time: 3 h

Plate: Pre-coated

Protocol: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to B-Raf Proto Oncogene Serine/Threonine Protein Kinase (BRAF). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to B-Raf Proto Oncogene Serine/Threonine Protein Kinase (BRAF). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain B-Raf Proto Oncogene Serine/Threonine Protein Kinase (BRAF), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of B-Raf Proto Oncogene Serine/Threonine Protein Kinase (BRAF) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Assay Precision: Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level B-Raf Proto Oncogene Serine/Threonine Protein Kinase (BRAF) were tested 20 times on one plate, respectively
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level B-Raf Proto Oncogene Serine/Threonine Protein Kinase (BRAF) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Restrictions: For Research Use only

Handling

Handling Advice: The Stop Solution is acidic. Do not allow to contact skin or eyes. Calibrators, controls and specimen samples should be assayed in duplicate. Once the procedure has been started, all steps should be completed without interruption.

Storage: 4 °C,-20 °C

Storage Comment: -20°C. Bring all reagents to room temperature before beginning test. The kit may be stored at 4°C for immediate use within two days upon arrival. Reseal any unused strips with desiccant pack. Minimize freeze/thaw cycles.

Expiry Date: 12 months

Target Details for BRAF

Target: BRAF (B-Raf proto-oncogene, serine/threonine kinase (BRAF))

Alternative Name: B-Raf Proto Oncogene Serine/Threonine Protein Kinase

Background: Gene Aliases: B-raf 1, BRAF1, RAFB1, V-Raf Murine Sarcoma Viral Oncogene Homolog B1

Gene ID: 673

UniProt: P15056

Pathways: MAPK Signaling, RTK Signaling, Neurotrophin Signaling Pathway, Ribonucleoprotein Complex Subunit Organization, Hepatitis C, Autophagy