Western blot abreview for Anti-Topoisomerase I antibody - ChIP Grade

Excellent
Abreviews
Application
Western blot
Sample
Human Cell lysate - other (top1 cleavage complex purify by CscL gradient)
Gel Running Conditions
Non-reduced Denaturing (Slot Blot)
Loading amount
2 µg
Treatment
DMSO or 10uM CPT
Specification
top1 cleavage complex purify by CscL gradient
Blocking step
Licor Blocking Buffer as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C

Other product details

Incubation time
2 hour(s) and 0 minute(s) · Temperature: 25°C
Dilution
1/1000

Secondary antibody

Dilution
1/10000
Name
Non-Abcam antibody was used: 800CW anti Rb (licor Odyssey)
Host species: Goat
Clonality: Polyclonal
Conjugation: IRDye® 800CW

Detection

Detection method
Odyssey CLx
Negative control
cells were treated with DMSO (line 1). Then, cells were lysed with 1% Sarkozyl (+ Protease inhibitors). Proteins covalently bind to DNA was purify by CsCl and load in slotblot.
Exposure
16 minute(s) and 0 second(s)
Bands
Specific: 1 kDa
Positive control
cells were treated with 10uM CPT (line 2). Then, cells were lysed with 1% Sarkozyl (+ Protease inhibitors). Proteins covalently bind to DNA was purify by CsCl and load in slotblot.

Additional data

Additional Notes
This experiment is not a classical western blot, since the material is load directly into a nitrocellulose membrane without passing through a acrylamide gel. Therefore, we don't separate the proteins by size.

Ms. Cristina Bernal

Verified customer

Submitted Mar 26 2019

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