Application
Western blot
Sample
Arabidopsis thaliana Recombinant protein (total seed extract)
Specification
total seed extract
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Other product details
Dilution
1/100
Incubation time
1 hour(s) and 0 minute(s)
Secondary antibody
Name
Non-Abcam antibody was used: Chemicon Aq330A ShXMs IgG F(ab')2
Host species: Sheep
Clonality: Polyclonal
Conjugation: Alkaline Phosphatase
Host species: Sheep
Clonality: Polyclonal
Conjugation: Alkaline Phosphatase
Detection
Detection method
NBT/BCIP
Exposure
1 minute(s) and 0 second(s)
Bands
Specific: 34.2 kDa
Positive control
Human insulin (non-reduced) [Sigma]
Negative control
Non recombinant Arabidopsis thaliana seed
Additional data
PubMed ID
not available yet: View
Additional Notes
The work was published in the Plant Biotechnology Journal Vol4:77-85 (2006)
The antibody worked well in the detection of fusion protein in transgenic seed. The antibody will weakly detect non-reduced insulin (needed to load at least 0.5 ug). Also detected reduced or non-reduced proinsulin weakly (need to load at 0.25 ug). Therefore it is difficult to use effectively in the detection of mature insulin or proinsulin, but has been extremely useful in the detection of recombinant fusion proteins with insulin-based polypeptides (authentic B-chain and A-chain primary amino acid sequence with variations of C-propeptide).
The antibody worked well in the detection of fusion protein in transgenic seed. The antibody will weakly detect non-reduced insulin (needed to load at least 0.5 ug). Also detected reduced or non-reduced proinsulin weakly (need to load at 0.25 ug). Therefore it is difficult to use effectively in the detection of mature insulin or proinsulin, but has been extremely useful in the detection of recombinant fusion proteins with insulin-based polypeptides (authentic B-chain and A-chain primary amino acid sequence with variations of C-propeptide).
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
Dr. Cory L Nykiforuk
Verified customer
投稿 Feb 24 2006