H2AFX ELISA Kit

Catalog No. ABIN6810525

This Colorimetric ELISA kit is designed for the quantitative measurement of Human, Mouse and Rat H2AFX.

Quick Overview for H2AFX ELISA Kit (ABIN6810525)

Target: H2AFX (H2A Histone Family, Member X (H2AFX))

Binding Specificity: acetylated

Reactivity: Human, Mouse, Rat

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Sample Type: Cell Lysate

Product Details H2AFX ELISA Kit

Purpose: Human, Mouse, Rat Acetylated-Histone H2AX ELISA Kit. This ELISA is for measuring Acetylated-Histone H2AX in Human, Mouse, Rat cell lysates.

Analytical Method: Semi-Quantitative

Specificity: The antibody pair provided in this kit recognizes Human///Mouse///Rat Acetylated-Histone H2AX.

Characteristics:

• Simultaneously measure cleaved protein and pan protein in one experiment (for normalization purpose)
• Screen numerous different cell lysates without performing a Western Blot analysis
• Minimal hands-on time, convenient, and non-radioactive material

Components:

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Anti-cleaved Antibody
• Anti-Pan Antibody
• HRP-Conjugated Secondary Antibody
• Streptavidin-Conjugated HRP
• Assay Diluent
• TMB One-Step Substrate
• Stop Solution
• Lysis Buffer
• Positive Control Sample

Material not included:

• Distilled or deionized water
• 100 mL and 1 liter graduated cylinders
• Tubes to prepare sample dilutions
• Protease and Phosphatase inhibitors
• Precision pipettes to deliver 2 μL to 1 mL volumes
• Adjustable 1-25 mL pipettes for reagent preparation
• Benchtop rocker or shaker
• Microplate reader capable of measuring absorbance at 450 nm

Application Details

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

Protocol:

1. Prepare all reagents and samples as instructed in the manual.
2. Add 100 μL of sample or positive control to each well.
3. Incubate 2.5 h at RT or O/N at 4 °C.
4. Add 100 μL of prepared primary antibody to each well.
5. Incubate 1 h at RT.
6. Add 100 μL of prepared 1X HRP-Streptavidin to each well.
7. Incubate 1 h at RT.
8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 μL of Stop Solution to each well.
11. Read at 450 nm immediately.

Restrictions: For Research Use only

Handling

Storage: -20 °C

Storage Comment: Upon receipt, the kit should be stored at -20 °C. Please use within 6 months from the date of shipment. After initial use, Wash Buffer Concentrate (Item B), Assay Diluent (Item E), TMB One-Step Substrate Reagent (Item H), HRP-Streptavidin (Item G), Stop Solution (Item I) and Cell Lysate Buffer (Item J) should be stored at 4 °C to avoid repeated freeze-thaw cycles. Return unused wells to the pouch containing desiccant pack, reseal along entire edge and store at -20 °C. Reconstituted Positive Control (Item K) should be stored at -70 °C.

Expiry Date: 6 months

Target Details for H2AFX

Target: H2AFX (H2A Histone Family, Member X (H2AFX))

Alternative Name: Histone H2AX

Gene ID: 3014

Pathways: Telomere Maintenance, DNA Damage Repair, Positive Regulation of Response to DNA Damage Stimulus