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 and Rat Acetylated-Histone H2AX ELISA Kit. This ELISA is for measuring Acetylated-Histone H2AX in human, mouse and rat cell lysates.

Analytical Method: Semi-Quantitative

Specificity: The antibody pair provided in this kit recognizes Human

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
• Assay Diluent
• Anti-Acetylated Antibody
• HRP-Conjugated Secondary Antibody
• TMB One-Step Substrate
• Stop Solution
• Lysis Buffer
• Positive Control Sample

,

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Assay Diluent
• Anti-Acetylated Antibody
• HRP-Conjugated Secondary Antibody
• 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.

Plate: Pre-coated

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 â€""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), 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 â€""20^°C. Item D, store at 2-8^°C for up to one month (store at -20^°C for up to 6 months, avoid repeated freeze-thaw cycles). 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

Background:

Gene Symbols: H2AFX, H2AX, H2A.X, HIST5-2AX

Background: Acetylated-Histone H2AX ELISA kit is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in human, mouse and rat cell lysates. By determining Acetylated Histone H2AX protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.

This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human, mouse and rat Acetylated-Histone H2AX. An anti-pan Histone H2AX antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and Histone H2AX present in a sample is bound to the wells by the immobilized antibody. The wells are washed and rabbit anti- Acetyl Lysine antibody is used to detect Acetylated Histone H2AX. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG 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 Acetylated Histone H2AX 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): H2a/x H2A.X

Gene ID: 3014

UniProt: P16104, P27661

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