Western blot abreview for Anti-gamma H2A.X (phospho S139) antibody [9F3]

Average
Abreviews
Application
Western blot
Sample
Mouse Recombinant protein (C-terminus of H2A.X with full range of phosphoryla)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (Dot blot)
Loading amount
0.384 µg
Specification
C-terminus of H2A.X with full range of phosphoryla
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Other product details

Incubation time
12 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: TBST with 5% skimmed milk
Dilution
1/2000

Secondary antibody

Dilution
1/10000
Name
Non-Abcam antibody was used: Bio-Rad Goat anti- mouse Cat no. 170-6516
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase

Detection

Detection method
ECL
Exposure
2 minute(s) and 30 second(s)

Additional data

Additional Notes
Figure legend: Mouse (left): 1: SQEY (non-phosphorylated SQEY motif of mouse H2A.X peptide), 2: pSQEY (serine monophosphorylated SQEY motif of mouse H2A.X peptide), 3: SQEpY (tyrosine monophosphorylated SQEY motif of mouse H2A.X peptide), 4: pSQEpY (serine and tyrosine di-phosphorylated SQEY motif of mouse H2A.X peptide). A: 384 ng (256 ng/uL), B: 96 ng (64 ng/uL), C: 24 ng (16 ng/uL), D: 6 (4 ng/uL), E: 1.5 (1 ng/uL) Conclusion: Little cross reactivity with other phosphorylation states observed with the analysed concentration of antibody. Human (right): 1: pSQEY (serine monophosphorylated SQEY motif of human H2A.X peptide), 2: pSQEpY (serine and tyrosine di-phosphorylated SQEY motif of human H2A.X peptide). A: 384 ng (256 ng/uL), B: 96 ng (64 ng/uL), C: 24 ng (16 ng/uL), D: 6 (4 ng/uL), E: 1.5 (1 ng/uL) Conclusion: Substantial cross reactivity with other phosphorylation states observed with the analysed concentration of antibody.
Username

Peter Ellis

Verified customer

Submitted Jun 02 2016

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