Western blot abreview for Anti-Histone H3 (phospho S10) [mAbcam 14955] antibody - ChIP Grade

Good
Abreviews
Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (10%)
Sample
Fruit fly (Drosophila melanogaster) Tissue lysate - whole (0-4hr embryos)
Specification
0-4hr embryos
Treatment
control shRNA and wee shRNA embryos
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C

Other product details

Incubation time
12 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% BSA/TBST
Dilution
1/1000

Secondary antibody

Name
Non-Abcam antibody was used: Amersham Anti-mouse IgG, peroxidase-linked whole a
Host species: Sheep
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/10000

Detection

Detection method
Pierce ECL
Bands
Specific: 15 kDa Non-specific: faint 45 kDa
Exposure
5 second(s)
Positive control
0-4hr old wee shRNA embryos (lane 2) should display elevated pHH3Ser10 levels relative to 0-4hr old EGFP shRNA embryos (lane 1)

Additional data

Additional Notes
lysis was in 8M urea by homogenization with Bullet blender (Next Advance) PVDF membrane for 1 hour transfer total HH3 blot below anti-HH3pSer10 blot to monitor loading

Dr. Richelle Sopko

Verified customer

Submitted Dec 18 2013

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