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

Poor
Abreviews
Application
Western blot
Sample
Zebrafish Tissue lysate - whole (4 dpf embryos (LWT))
Gel Running Conditions
Reduced Denaturing (4-15 % gradient)
Loading amount
60 µg
Treatment
DMSO 2 h
Specification
4 dpf embryos (LWT)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Other product details

Incubation time
18 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: TBST
Dilution
1/500

Secondary antibody

Dilution
1/4000
Name
Non-Abcam antibody was used: IgG (H + L)-HRP Conjugate
Host species: Rabbit
Clonality: Polyclonal
Conjugation: HRP polymer

Detection

Detection method
ECL
Exposure
5 minute(s) and 0 second(s)
Positive control
60 ug HEK293 whole cell lysate

Additional data

Additional Notes
Nitrocellulose membrane (Bio-Rad, 170-4271) Lysis buffer: 200 mM Hepes, 40 mM NaCl, 2mM MgCl2, 0.5 % Triton X-100, 1 x protease inhibitor cocktail (Roche, 4693159001), 1 x phosphatase inhibitor cocktail (Roche, 4906837001), 25 U/mL BaseMuncher (Expedeon, BM0025) Lanes: 1 - zebrafish lysate 2 - HEK293 lysate 250 - 200 kDa anti-ATM 200 - 50 anti - TOP1 50 - 10 anti-RNase H2a Blot was reprobed for tubulin and actin loading controls ~50 kDA: tubulin 1:1000 dilution actin 1:1000 dilution
Abcam response
Abcam had not tested the antibody in this Zebrafish samples and thus can't say how it should have performed. This individual received a credit for testing the antibody in Zebrafish samples as part of our AbTrial program.

Dr. Ringaile Zaksauskaite

Verified customer

Submitted Sep 07 2018

Sign up