Western blot abreview for Anti-Nestin antibody [SP103]

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Abreviews
Application
Western blot
Loading amount
5 µg
Gel Running Conditions
Reduced Denaturing (10%)
Sample
Human Cell lysate - other (undifferentiated human iPS cells)
Specification
undifferentiated human iPS cells
Blocking step
5% milk in TBST 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) · Diluent: 5% milk in TBS
Dilution
1/100

Secondary antibody

Name
Non-Abcam antibody was used: Dako Goat anti-Rabbit IgG
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/2000

Detection

Detection method
ECL
Bands
Specific: ~178 kDa
Exposure
5 minute(s) and 0 second(s)

Additional data

Additional Notes
Undifferentiated human iPS cells were lysed in RIPA buffer with protease inhibitors and performed under reducing conditions (10% gel). Following block overnight the samples were probed using an anti Nestin antibody (ab105389) at 1ug/ml, followed by incubation with the relevant secondary antibody at 1:2000. Samples were visualised using ECL for 5 mins and a single band at approx 178kDa was observed. Although undifferentiated cells should not express Nestin, there are obviously some cells within the sample that have begun differentiation.

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Submitted Dec 18 2013

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