Western blot abreview for Anti-Actin antibody [2Q1055]

Good
Abreviews
Application
Western blot
Sample
Chlamydomonas reinhardtii Cell lysate - whole cell (Chlamydomonas reinhardtii)
Loading amount
20 µg
Specification
Chlamydomonas reinhardtii
Treatment
Various concentrations of calcium (0M to 10^-3M)
Gel Running Conditions
Non-reduced Denaturing (12% gradient gel (polyacrylamide))
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1.5% · Temperature: 23°C

Other product details

Dilution
1/1000
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: TBST with 1.5% dry milk

Secondary antibody

Name
Non-Abcam antibody was used: Santa Cruz Biotechnology goat anti-mouse IgG-AP
Host species: Goat
Clonality: Polyclonal
Conjugation: Alkaline Phosphatase
Dilution
1/5000

Detection

Detection method
NBT/BCIP
Exposure
45 minute(s) and 0 second(s)
Bands
Specific: 43 kDa
Positive control
I did not use a positive control for this blot.

Additional data

Additional Notes
This product has never been tested for reactivity in Chlamydomonas reinhardtii. The depicted blot was blocked with 1.5% dry milk and the antibody was in TBST with 1.5% dry milk. I have a test blot (half blot) with a 1% block and primary in TBST with 1% dry milk that has stronger bands. I also developed the blot longer to see if the bands would be more robust but they were not. Bands appeared at around 15 minutes.

lanes are as follows:
1. SeeBlue® Plus2 Pre-Stained Standard
2. No calcium
3. 10^-7M free calcium
4. 10^-6M free calcium
5. 10^-5M free calcium
6. 10^-4M free calcium
7. 10^-3M free calcium

Caroline Lam

Verified customer

Submitted May 10 2013

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