Western blot Abreview for CARHSP1 protein

Good
Abreviews
Application
Western blot

Review text: We used this HA-tagged CARHSP1 protein only as a control to quantify our own HA-tagged proteins on a Western blot. For this it worked well, although there are weak degradation products as well as the main 44 kDa band for CARHSP1.
Sample: Human Recombinant protein
Loading amount: 0.3 µg
Treatment: diluted in 0.8M Tris, 0.2M sorbitol, 1% betamercaptoethanol, with bromophenol blue. Added 1% SDS then boiled for 5 min.
Gel Running Conditions: Reduced Denaturing (15% acrylamide gel)
Blocking step: Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 10°C

Other product details
Dilution: 1/100
Incubation time: 1 hour(s) and 0 minute(s) · Temperature: 20°C · Diluent: TBS + 0.1% tween

Primary antibody (in addition to 'CARHSP1 protein')
Name: Non-Abcam antibody was used: anti-HA
Host species: Rabbit
Dilution: 1/2000

Secondary antibody
Name: Non-Abcam antibody was used: anti-rabbit ECL
Clonality: Polyclonal
Dilution: 1/5000

Detection
Detection method: ECL
Exposure: 4 second(s)
Bands: Specific: 44 kDa Non-specific: 30, 35 kDa

Additional data
Additional Notes: Non-specific bands (30, 35kDa) may be due to degradation.

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Submitted Sep 24 2012

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