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

Average
Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (SCLC xenograft, NSCLC, A431, RH30 cells)
Loading amount
20 µg
Specification
SCLC xenograft, NSCLC, A431, RH30 cells
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

Other product details

Dilution
1/5000
Incubation time
1 hour(s) and 0 minute(s) · Diluent: TBS/5% NFDM

Secondary antibody

Name
Non-Abcam antibody was used: Anti-Rb IgG (H&L)-HRP, Cell Signalling #7074
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/25000

Detection

Detection method
SuperSignal West Femto
Exposure
30 second(s)
Bands
Specific: 92 kDa Non-specific: 35, 21 kDa
Positive control
SW210.5 (SCLC) xenograft lysate and 5M2 (NSCLC), A431, Rh30 cell lysates
Negative control
normal lung tissue lysate

Additional data

Additional Notes
It is possible that the non-specific band at ~35 kDa may be a fragment of the TOPO I since the smaller band appears stronger when the larger band is lighter. I haven't checked this in detail, however.

The WB was performed using Millipore PVDF.

20 ug protein/lane.

Lane assignments:
1. SW210.5 human SCLC cell xenograft lysate.
2. 5M2 human NSCLC cell lysate (bicarb. buffer).
3. 5M2 human NSCLC cell lysate (M-Per buffer).
4. Normal human lung tissue lysate.
5. A431 cell (human epidermoid carcinoma) lysate .
6. Rh30 cell (human rhabdomyosarcoma) lysate.

Dr. Mike Campa

Verified customer

Submitted Jun 27 2007

Sign up