Application
Western blot
Sample
Schizosaccharomyces pombe Purified protein (Purified modified histone peptides)
Loading amount
0.1 µg
Specification
Purified modified histone peptides
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Other product details
Dilution
1/1000
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% Milk PBST
Secondary antibody
Name
Non-Abcam antibody was used: Anti-rabbit
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/1000
Detection
Detection method
ECL
Exposure
15 second(s)
Bands
Specific: 0 kDa
Positive control
I used purified peptides of modified histone H4. I used Histone H4 lysine (K) 5-acetylated (H4K5ac),
Negative control
I used unmodified H4, H4K4K8ac and H4K12ac.
Additional data
Additional Notes
The method used was a slot blot. I used purified proteins to test specificity. Compared to the current anti-H4K5ac antibody used in lab, Abcam's antibody gave a higher background and less H4K5ac specificity.
The picture shows two antibodies against purified peptides, the Abcam blot is labeled Abcam. There are 4 vertical lanes and 3 horizontal rows. The 4 vertical lanes correspond to the peptides used. Lane 1: unmodified histone H4, Lane 2: H4K5ac, Lane 3: H4K8ac, Lane 4: H4K12ac. The horizontal rows indicate concentration. Row 1 is 100 ng/ul and each subsequent row is a 1:10 dilute of the previous spot.
The picture shows two antibodies against purified peptides, the Abcam blot is labeled Abcam. There are 4 vertical lanes and 3 horizontal rows. The 4 vertical lanes correspond to the peptides used. Lane 1: unmodified histone H4, Lane 2: H4K5ac, Lane 3: H4K8ac, Lane 4: H4K12ac. The horizontal rows indicate concentration. Row 1 is 100 ng/ul and each subsequent row is a 1:10 dilute of the previous spot.
Abcam response
Many thanks for your honest and valuable feedback. Unfortunately, S. pombe is not a tested species for ab51997 and your data suggests it may not be suitable for this species under the conditions in which you tested. Thank you for your time - we hope that this information will be helpful for future customers.
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.
Abcam user community
Verified customer
投稿 Feb 03 2010