H2AFX ELISA Kit

Catalog No. ABIN2748159

Product Details H2AFX ELISA Kit

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

Binding Specificity: pSer139, total

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Purpose: Human Phospho-H2AX (S139) and Total H2AX ELISA Kit. This assay semi-quantitatively measures phophorylated H2AX (Ser139) and Total H2AX in lysate samples.

Sample Type: Cell Lysate, Tissue Lysate

Analytical Method: Semi-Quantitative

Specificity: The antibody pair provided in this kit recognizes human H2AX phosphorylated at Serine-139 and total H2AX.

Characteristics:

• Simultaneously measure Phosphorylated 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-Phospho 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

Sample Volume: 100 μL

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.

Assay Procedure:

Prepare all reagents and samples as instructed in the manual.
Add 100 μL of sample or positive control to each well.
Incubate 2.5 h at RT or O/N at 4 °C.
Add 100 μL of prepared primary antibody to each well.
Incubate 1 h at RT.
Add 100 μL of prepared 1X HRP-Streptavidin to each well.
Incubate 1 h at RT.
Add 100 μL of TMB One-Step Substrate Reagent to each well.
Incubate 30 min at RT.
Add 50 μL of Stop Solution to each well.
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: H2AX (H2AFX Products)

Synonyms: H2A.X ELISA Kit, H2A/X ELISA Kit, H2AX ELISA Kit, AW228881 ELISA Kit, H2ax ELISA Kit, Hist5-2ax ELISA Kit, gammaH2ax ELISA Kit, zgc:56329 ELISA Kit, h2a.x ELISA Kit, h2a/x ELISA Kit, h2ax ELISA Kit, RGD1566119 ELISA Kit, h2a ELISA Kit, h2afx ELISA Kit, H2A histone family member X ELISA Kit, H2A histone family, member X ELISA Kit, H2A histone family member X L homeolog ELISA Kit, histone cluster 1, H2ah ELISA Kit, histone cluster 2, H2ab S homeolog ELISA Kit, histone H2AX ELISA Kit, H2AFX ELISA Kit, H2afx ELISA Kit, h2afx ELISA Kit, h2afx.L ELISA Kit, HIST1H2AH ELISA Kit, hist2h2ab.S ELISA Kit, LOC100522201 ELISA Kit, LOC100720536 ELISA Kit

Background: Histone H2AX phosphorylated at Serine-S139 and total H2AX

Gene ID: 3014

UniProt: P16104

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