A rabbit polyclonal antibody against a 242 amino acid sequence in the N-terminus of X. laevis TPX2 was raised by Covance and affinity purified from total serum on a HiTrap N-hydroxysuccinimide–activated HP column (GE Healthcare) coupled with recombinant full length X. laevis TPX2. Antibodies were eluted with Gentle Ag/Ab Elution Buffer (Thermo Fisher Scientific) and dialyzed into 50 mM Hepes. The 242 amino acid sequence is highly conserved between X. laevis and X. tropicalis (87% identical) and X. laevis and H. boettgeri (86% identical), and was used at a 1:5,000 dilution for Western blot, and a 1:5,000 dilution for immunofluorescence.
Hitrap n hydroxysuccinimide activated hp column
Manufactured by GE Healthcare
The HiTrap N-hydroxysuccinimide–activated HP column is a prepacked affinity chromatography column designed for the immobilization and purification of proteins. The column matrix is composed of agarose beads with N-hydroxysuccinimide (NHS) groups that can covalently bind to primary amine groups on proteins. This allows for the efficient capture and separation of target proteins from complex mixtures.
Automatically generated - may contain errors
2 protocols using hitrap n hydroxysuccinimide activated hp column
1
Affinity Purification of X. laevis TPX2 Antibody
2
Immunodepletion and Reconstitution of Nap1 in Xenopus Egg Extracts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Rabbit polyclonal antibodies against full-length Nap1 were raised by Covance and affinity purified from total serum on a HiTrap N-hydroxysuccinimide–activated HP column (GE Healthcare) coupled with recombinant Nap1. Antibodies were eluted with Gentle Ag/Ab Elution Buffer (Thermo Fisher Scientific) and dialyzed into 50 mM Hepes.
H1M was immunodepleted from egg extracts as previously described (Maresca et al., 2005 (link)). In brief, 55 µl CSF extract was subjected to two successive 45-min incubations with 40 µg anti-H1M antibody coupled to 200 µl protein A Dynabeads (Invitrogen). To deplete Nap1, 11 µg of affinity-purified Nap1 antibody coupled to 40 µl protein A Dynabeads (Invitrogen) was used to deplete 11 µl of egg extract over two rounds of 45-min incubations. For control (mock depleted) reactions, an equal amount of total rabbit IgG antibody was coupled to beads. Recombinant Nap1-L1B or mutants (Nap1N6, Nap1C9, and Nap1N6C9) were added back to depleted extracts at a final concentration of 1 µM to match endogenous Nap1 levels.
H1M was immunodepleted from egg extracts as previously described (Maresca et al., 2005 (link)). In brief, 55 µl CSF extract was subjected to two successive 45-min incubations with 40 µg anti-H1M antibody coupled to 200 µl protein A Dynabeads (Invitrogen). To deplete Nap1, 11 µg of affinity-purified Nap1 antibody coupled to 40 µl protein A Dynabeads (Invitrogen) was used to deplete 11 µl of egg extract over two rounds of 45-min incubations. For control (mock depleted) reactions, an equal amount of total rabbit IgG antibody was coupled to beads. Recombinant Nap1-L1B or mutants (Nap1N6, Nap1C9, and Nap1N6C9) were added back to depleted extracts at a final concentration of 1 µM to match endogenous Nap1 levels.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!