Western blot abreview for Anti-PDGF Receptor alpha (phospho Y849) antibody

Below Average
Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (Brain Glioblastoma)
Loading amount
120 µg
Specification
Brain Glioblastoma
Treatment
PDGF 20/40 ng/ml 5, 10, 15 min
Gel Running Conditions
Reduced Denaturing (7,5 %)
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5%

Other product details

Dilution
1/500
Incubation time
12 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: and RT

Secondary antibody

Name
Non-Abcam antibody was used: mouse anti rabbit
Host species: Mouse
Clonality: Monoclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/10000

Detection

Detection method
ECL+
Exposure
10 minute(s) and 0 second(s)
Bands
Specific: 120 ? kDa Non-specific: 80, 60, 40, 25 kDa
Positive control
D54, U343 cells treated with PDGF
Negative control
untreated D54, U343 cells

Additional data

Additional Notes
Modifications tested:

PRIMARY ANTIBODY
ab 79318 (1:1000, 1:500 1:300 1:100) used (4°C and RT for 1h - overnight)

blocking with BSA 2%, 5%, 7,5 %, 10%
blocking with PhosphoBlocker 1 - 5%
blocking with Milk Powder 5% 10%
blocking with Slimfast 0,5%, 1%
each blocking step tested with RT and/or 4°C (1h - overnight)

secondary antibody concentration from 1:2500 1:5000 1:7500 1:10000 1:20.000

secondary antibody conjugation (HRP Biotin)

washing time and buffer (TBST TST)
Abcam response
We are sorry that this antibody has produced poor results and have credited the antibody to the customer.

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Submitted Oct 08 2010

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