Western blot abreview for Anti-beta Actin antibody [mAbcam 8224] - Loading Control

Excellent
Abreviews
Application
Western blot
Sample
Fruit fly (Drosophila melanogaster) Tissue lysate - whole (Whole cell lysate)
Loading amount
100 µg
Specification
Whole cell lysate
Gel Running Conditions
Reduced Denaturing (Novex 10% Bis-Tris)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Other product details

Dilution
1/1000
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 20°C · Diluent: 2.5% milk

Secondary antibody

Name
Non-Abcam antibody was used: ECL Sheep Anti-mouse IgG, HRP-linked (GE Healthcar
Host species: Sheep
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/10000

Detection

Detection method
Amersham ECL
Exposure
2 minute(s) and 0 second(s)
Bands
Specific: 42 kDa
Positive control
No
Negative control
No

Additional data

PubMed ID
19409408: View
Additional Notes
I like this antibodies. I had a problem with detection of Drosophila melanogaster beta-actin using monoclonal anti-beta-actin antibodies from Sigma (it is declared that Sigma antibodies can detect Drosophila beta-actin. Not in my hands((( ). When I have found mAbcam 8224 antibodies I was happy to add a good panel to the figure in my Drosophila paper. Flies were homogenized in liquid nitrogen, resuspended in PBS with protease inhibitor mixture (Roche), incubated on ice for 20 minutes and centrifuged at max speed for 15 minutes. Supernatant was used for WBs. 100 ug of total protein per lane was loaded. I used this antibody for mouse tissues and cell culture too, they always produced paper quality pictures. I strongly recommend these Abs for WBs with Drosophila homogenates.

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Submitted Mar 05 2010

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