Application
Western blot
Sample
Mouse Purified protein (Liver)
Gel Running Conditions
Reduced Denaturing (4-12)
Loading amount
0.0001 µg
Specification
Liver
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 1%
Other product details
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 1% Skim Milk in TBST
Dilution
1/2000
Secondary antibody
Dilution
1/5000
Name
Non-Abcam antibody was used: Amersham ECL HRP Conjugated Antibodies
Host species: Rabbit
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Rabbit
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL
Negative control
Empty lane X (loading buffer without protein)
Exposure
4 minute(s) and 0 second(s)
Bands
Specific: 68 kDa
Positive control
Protein complex containing Cryptochrome-1, NativeMark Unstained Protein Standard (Thermo Fischer Scientific)
Additional data
Additional Notes
Blue Native PAGE:
Protein complex containing Cryptochrome-1 was purified and loaded onto a glycerol gradient. Fractions were taken [1-7] and loaded onto a Blue Native PAGE (4-16% Bis-Tris) gel along with an empty lane as a control, and NativeMark Unstained Protein Standard. Electrophoresis for 2 hours, transfer onto PVDF membrane for 1 hour. Block in skim milk (1%) at room temperature. Primary antibody incubation for 1 hour at room temperature, followed by secondary incubation. Blots were developed using ECL solution.
SDS-PAGE
Protein complex containing Cryptochrome-1 was purified. The purified sample was split and incubated with Protein G Dynabeads bound to various antibodies, including anti-Cryptochrome 1 (ab104736). Immunodepletion by anti- Cryptochrome-1 antibody was successful in depleting Cryptochrome 1 bound protein complex from the sample.
Band at ~ 68 kDa on SDS-PAGE
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
Mariska Twaalfhoven
Verified customer
提交于 Dec 15 2015