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H2AFX ELISA Kit (Human)

CAT: 0247-OKCD02089-96WSize: 96 WellDry Ice: NoHazardous: No
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CAT#:0247-OKCD02089-96WSize:96 Well
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Reproducibility
Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level H2A Histone Family, Member X (H2AFX) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level H2A Histone Family, Member X (H2AFX) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%
CAS Number
7732-18-5
Gene Name
H2A.X variant histone
Gene Aliases
H2A histone family member X; H2A.X; H2A/X; H2AFX; H2AX histone; histone H2A.x; histone H2AX.
Gene ID
3014
Accession Number
NP_002096.1
Reactivity
Homo sapiens|Human
Target
Variant histone H2A which replaces conventional H2A in a subset of nucleosomes. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. Required for checkpoint-mediated arrest of cell cycle progression in response to low doses of ionizing radiation and for efficient repair of DNA double strand breaks (DSBs) specifically when modified by C-terminal phosphorylation.
Type
ELISA Kit
Applications
Enzyme-linked Immunosorbent assay-Sandwich
Detection Method
Colorimetric
Assay Principle
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 H2A Histone Family, Member X (H2AFX) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to H2A Histone Family, Member X (H2AFX) . 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 H2A Histone Family, Member X (H2AFX), 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 H2A Histone Family, Member X (H2AFX) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Assay Protocol
Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol
Sample Type
Tissue homogenates, cell lysates cell culture supernatants and other biological fluids
Detection Range
0.156-10ng/mL
Sensitivity
< 0.059 ng/mL
Reconstitution
2°C to 8°C|-20°C
Components
Component Amount Anti-H2AFX Microplate 96 Wells (12 x 8 Well Strips) H2AFX Lyophilized Standard 2 x 10 ng 100X Biotinylated H2AFX Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Component Amount Component Amount Anti-H2AFX Microplate 96 Wells (12 x 8 Well Strips) Anti-H2AFX Microplate 96 Wells (12 x 8 Well Strips) H2AFX Lyophilized Standard 2 x 10 ng H2AFX Lyophilized Standard 2 x 10 ng 100X Biotinylated H2AFX Detector Antibody 120 uL 100X Biotinylated H2AFX Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL
Notes
Formerly GWB-KBBGN3
Specificity
This assay has high sensitivity and excellent specificity for detection of H2A Histone Family, Member X (H2AFX) . No significant cross-reactivity or interference between H2A Histone Family, Member X (H2AFX) and analogues was observed.
Protein Length
143
NCBI Gene Symbol
H2AX
Protein Name
Histone H2AX
Gene Name URL
H2AX
Nucleotide Accession Number
NM_002105.2

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