Result“Breast and lung cancer cell lines with various treatments were tested for this antibody. With regular ECL a band was barely detected at 20 minutes. With super signal pretty clean bands were obtained.” – External Laboratory

Antibody Customer Review: STJ97186 Anti-Histone H2A.X (Phospho-Ser139) Antibody

Antibody Specificity: Antibody Rating:

ProtocolTreatment of materials: 5 Gy radiation.

E.g. Gel electrophoresis: SDS gel electrophoresis.

Transfer: Wet transfer.

Blocking: 3% milk for 30 minutes.

Membrane wash: 3X 5 minutes after primary and secondary antibodies with PBST.

Primary antibody probing: 1:500 dilution at 4 degree Celsius overnight.

Membrane wash: 3X 5minutes after primary and secondary antibodies with PBST.

Secondary antibody probing: 1 hour at room temperature (1:2500 dilution).

Membrane wash: 3X 5minutes after primary and secondary antibodies with PBST.

Visualization: ECL visualization for 5 seconds with super signal.

Cell Line: Breast and Lung cancer cell lines

Method of validation: Western Blot

Primary Antibody: STJ97186 Anti-Histone H2A.X (Phospho-Ser139) Antibody

Secondary Antibody Goat anti-rabbitReference Antibody Beta ActinDilution ratio: 1:500

Gel electrophoresis information: SDS gel Electrophoresis

Transfer information: Wet Transfer.

Lane No. AntigenLoading amount

Primary antibody

Primary antibody

dilution ratio

Secondaryantibody dilution

ratioTarget band KD

Visualization time

1 MDA-MB-468 25ug STJ97186 Anti-Histone

H2A.X (Phospho-Ser139)

Antibody

1:500 1:2500 ~14kDa5 seconds with super

signal5 H1299 18ug

9 DCIS 25ug

The figure provided shows the bands obtained from MDA-MB-468, H1299 and DCIS cells. Only the control cells were indicated in the names because of the confidentiality. The bands appear to be specific. An incresae in the protein was detected in lane 4 and lane 8 as expected.