Western blot abreview for Anti-SEPT7 antibody

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Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa cells)
Gel Running Conditions
Reduced Denaturing (4-12% gradient)
Loading amount
10 µg
Specification
HeLa cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Other product details

Incubation time
20 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% milk PBST
Dilution
1/1000

Secondary antibody

Dilution
1/5000
Name
Non-Abcam antibody was used: AffiniPure Donkey Anti-Rabbit IgG (H+L) (jaackson
Host species: Rabbit
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase

Detection

Detection method
ECL prime
Exposure
5 minute(s) and 0 second(s)
Bands
Specific: 50 kDa

Additional data

Additional Notes
Lane 1 10ug of HeLa cell lysate loaded on 4-12% bis tris gel run in 1X MOPS buffer blot probed with primary septin 7 antibody ab186010 incubated 1:1000. lysis buffer 1.5X lammeli buffer 0.15M DTT WB protocol Western blot was performed by wet transfer of gel onto nitrocellulose membrane 100V for 2 h 4 degrees. Blocked in 5% milk PBST for 1 h room temp (1X PBS + 0.1% tween 20) incubated 24h at 4 degrees in 5% PBST with primary antibody 1:1000 Washed 3X PBST c IgG anti rabbit HRP 1:5000 (Jackson Immunoscience) Washed 3x PBST Washed 1X PBS Developed using ECL prime kit (GE healthcare)

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Submitted Nov 07 2018

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