Western blot abreview for Anti-SOX9 antibody

Average
Abreviews
Application
Western blot
Loading amount
30000 cells
Gel Running Conditions
Reduced Denaturing (11)
Sample
Human Cell lysate - whole cell (MCF-7 cells (human breast adenocarcinoma cell line)
Specification
MCF-7 cells (human breast adenocarcinoma cell line
Treatment
0.5% oxygen for 16hrs
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Other product details

Incubation time
1 hour(s) and 0 minute(s) · Temperature: 25°C · Diluent: PBS-Tween
Dilution
1/500

Secondary antibody

Name
Non-Abcam antibody was used: Polyclonal Swine anti-rabbit IgG/HRP
Host species: Pig
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/1000

Detection

Detection method
SuperSignal West Dura Chemiluminescent S
Bands
Specific: 55 kDa Non-specific: 30, 45, 60, 72, 80, kDa
Exposure
10 second(s)
Positive control
Hypoxic sample (treated with 0.5% oxygen for 16h). Predicted to be induced compared to normoxic sample based on mRNA analysis.

Additional data

Additional Notes
Lane 1 and 3 = MCF-7 (16h 0.5% O2) Lane 2 and 4 = MCF-7 (Normoxia) Lysis buffer: Urea/SDS buffer PVDF membrane used Comments: Bands detected at the right size but no clear induction observed in hypoxic sample. There are multiple non-specific bands observed.

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Submitted Nov 05 2013

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