My question pertains to the generation of polyclonal antibodies from synthetic peptides, such as those provided by GenScript, for use in ChIP-Seq. I recognize a potential problem in submitting a single peptide for antibody generation: that the specific protein epitope recognized by the antibody and represented by the synthetic peptide will not be exposed after formaldehyde cross-linking of the protein-DNA complex. Would it therefore be possible to obtain a polyclonal antibody by inoculating rabbits with two synthetic peptides from the same protein, such that more than one epitope may be recognized by the antibody?
Seqanswers Leaderboard Ad
Collapse
Announcement
Collapse
No announcement yet.
X
-
I don't think the 2-peptide strategy will work because each of the peptide will be independently displayed by the immune system. I know of a few companies that are trying this approach (with a single peptide) and are having marginal success.
If you are married to using a peptide antigen, you may want to try raising a monoclonal so you can screen for the desired properties (recognizing DNA-bound protein). Here though, the major challenge is having target to use in your screening.
Another thought would be to try a genetic immunization approach (like SDI). I've never tried them, but it looks look on paper...
Latest Articles
Collapse
-
by seqadmin
Proteins are often described as the workhorses of the cell, and identifying their sequences is key to understanding their role in biological processes and disease. Currently, the most common technique used to determine protein sequences is mass spectrometry. While still a valuable tool, mass spectrometry faces several limitations and requires a highly experienced scientist familiar with the equipment to operate it. Additionally, other proteomic methods, like affinity assays, are constrained...-
Channel: Articles
04-04-2024, 04:25 PM -
-
by seqadmin
Despite advancements in sequencing platforms and related sample preparation technologies, certain sample types continue to present significant challenges that can compromise sequencing results. Pedro Echave, Senior Manager of the Global Business Segment at Revvity, explained that the success of a sequencing experiment ultimately depends on the amount and integrity of the nucleic acid template (RNA or DNA) obtained from a sample. “The better the quality of the nucleic acid isolated...-
Channel: Articles
03-22-2024, 06:39 AM -
ad_right_rmr
Collapse
News
Collapse
Topics | Statistics | Last Post | ||
---|---|---|---|---|
Started by seqadmin, 04-11-2024, 12:08 PM
|
0 responses
25 views
0 likes
|
Last Post
by seqadmin
04-11-2024, 12:08 PM
|
||
Started by seqadmin, 04-10-2024, 10:19 PM
|
0 responses
27 views
0 likes
|
Last Post
by seqadmin
04-10-2024, 10:19 PM
|
||
Started by seqadmin, 04-10-2024, 09:21 AM
|
0 responses
24 views
0 likes
|
Last Post
by seqadmin
04-10-2024, 09:21 AM
|
||
Started by seqadmin, 04-04-2024, 09:00 AM
|
0 responses
52 views
0 likes
|
Last Post
by seqadmin
04-04-2024, 09:00 AM
|
Comment