Western blot abreview for Anti-TRPS1 antibody

Good
Abreviews
Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing (8%)
Sample
Mouse Cell lysate - nuclear (EpH4 (Non tumorogenic mammary epithelial cell line)
Specification
EpH4 (Non tumorogenic mammary epithelial cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C

Other product details

Incubation time
18 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: PBS-T+10%FCS
Dilution
1/500

Secondary antibody

Name
Non-Abcam antibody was used: ECL Rabbit IgG, HRP-linked whole Ab (from donkey),
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/10000

Detection

Detection method
Luminata Crescendo, Millipore WBLUR0500
Bands
Specific: 142 kDa Non-specific: 60-65, 85, 120 kDa
Exposure
20 second(s)
Negative control
EpF-1 and EpF-2 (EpH4 subclones expressing high amounts of Zeb1 and Zeb2 mRNA): According to qPCR data, TRPS1 mRNA levels are decreased in these samples. When compared to housekeeping, TRPS1 protein levels are decreased in these samples in the WB shown.
Positive control
EpH4 (parental cell line) were TRPS1 is expressed

Additional data

Additional Notes
Figure legend: 1.Ha-Ras-transformed EpH4 cells 2. and 3. EpF-2 and EpF-1 (EpH4 subclones expressing high amounts of Zeb1 and Zeb2 mRNA) 4.EpH4 (parental) 5.EpF-2 6. and 7.EpF-Z1.1 and EpF-Z1.2 (EpF-1 clones transfected with siRNA to knockdown Zeb1) 8. and 9. EpF-Z2.1 and EpF-Z2.2 (EpF-1 clones transfected with siRNA to knockdown Zeb2). Nuclear extracts were prepared in the presence of protein inhibitors and 25ug of protein were run in a 8% gel and transferred to a nitrocellulose membrane (transferred in 1xtransfer buffer+20% MeOH, 1.5h at 110v constant).

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Submitted Jan 28 2014

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