Western blot abreview for Anti-Tau (phospho S199) antibody [EPR2401Y]

Good
Abreviews
Application
Western blot
Loading amount
10 µg
Gel Running Conditions
Reduced Denaturing (10)
Sample
Human Cell lysate - whole cell (human stem cell derived neurons (healthy/diseased))
Specification
human stem cell derived neurons (healthy/diseased)
Treatment
quinolinic acid 72h (and non treated)
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: rt°C

Other product details

Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% BSA in TBST
Dilution
1/5000

Secondary antibody

Name
Non-Abcam antibody was used: DAKO goat anti rabbit IgG HRP conjugated
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/2000

Detection

Detection method
ECL
Bands
Specific: 55 kDa Non-specific: 100 kDa
Exposure
4 minute(s) and 0 second(s)

Additional data

Additional Notes
Human stem cell derived neurons were cultured for 30 days prior to collection and lysed in RIPA buffer with protease inhibitors. Proteins were resolved under reducing conditions (10% gel). Following block (5% BSA in TBST), the samples were probed using an anti phospho tau pSer199 antibody at 1:5000 in blocking buffer overnight at 4C. This was followed by incubation with the relevant secondary antibody at 1:2000. Samples were visualised using ECL for 4 mins and a specific band at approximately 55kDa was observed. However, a weaker non-specific band was also observed at 100kDa, this may be non-specific binding or potentially other isoforms. D = diseased cells (PSEN2 mutant) H = healthy cells.

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Submitted Mar 26 2014

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