Western blot abreview for Anti-Vinculin (phospho Y100) antibody

Inconclusive
Abreviews
Application
Western blot
Sample
Pig Cell lysate - whole cell (Ocular cell)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
Ocular cell
Blocking step
LICOR blocking buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

Other product details

Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5%BSA TTBS
Dilution
1/1000

Secondary antibody

Dilution
1/20000
Name
Non-Abcam antibody was used: GAR Alexa 790 conjugated secondary
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor­ 790

Detection

Detection method
LICOR odyssey
Negative control
None
Exposure
1 minute(s) and 0 second(s)
Bands
Specific: ~100 kDa
Positive control
None

Additional data

Additional Notes
abtrial TVTYI Ab200812 produced a band around the expected molecular weight (left). However, this band migrated faster (appeared as lower MW) than total vinculin protein detected by a separate antibody (middle, right). As phosphoprotein bands usually migrate slower than the total protein bands, it is inconclusive if AB200812 is detecting PY100-vinculin in pigs.
Abcam response
Thank you for submitting your feedback on use of ab200812 in pig samples. We have not validated this antibody in pig, and will continue to collect data on use in this species.

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Submitted Apr 14 2017

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