A complex of GnRH genes consisting of six copies of GnRH‐I and GnRH‐II (GnRH‐I+II) was generated. GnRH genes were conjugated with the gene encoding the STF‐2 protein (GenBank accession no. ERO08334) using a Gly‐Gly‐Gly‐Ser (GGGS) linker between its Ile 239 and Thr 243 residues. The synthesized genes were inserted into the pQE‐40 vector (Qiagen, Germany) by cleaving the BamHI and HindIII sites with restriction enzymes (New England Biolabs, USA). Recombinant GnRH proteins were expressed in Escherichia coli according to the manufacturer's instructions (Qiagen) (Hu et al., 2017 (link)). The identities of the recombinant proteins were determined by western blotting using rabbit anti‐GnRH (cat no. ab8491; Abcam, UK) and anti‐flagellin polyclonal antibodies (cat no. ab93713; Abcam), as previously described (Kim & Myoung, 2018 (link); Song et al., 2012 (link)).
Rabbit anti gnrh cat no ab8491
Manufactured by Abcam
Sourced in United Kingdom
Rabbit anti-GnRH (cat no. ab8491) is a primary antibody that recognizes the Gonadotropin-Releasing Hormone (GnRH) protein. It is produced in rabbit and can be used in various immunodetection techniques.
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2 protocols using rabbit anti gnrh cat no ab8491
1
Recombinant GnRH Protein Production
2
Generation of Recombinant GnRH Proteins
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Three types of recombinant GnRH proteins were generated as follows: 12 copies of GnRH-I, 12 copies of GnRH-II, and a complex consisting of 6 copies each of GnRH-I and -II (GnRH-I+II). All recombinant proteins were conjugated to STF-2 protein (GenBank accession no. ERO08334) by a GGGS linker between its isoleucine 239 and threonine 243 residues. The synthesized genes were inserted into the pQE-40 vector (Qiagen, Germany) by cleaving at BamHI and HindIII sites with restriction enzymes (New England Biolabs, USA). The recombinant GnRH proteins were expressed in E. coli according to the manufacturer's procedures (Qiagen) [11, 12] . The identities of recombinant proteins were determined by western blotting with rabbit anti-GnRH (cat no. ab8491; Abcam, UK) and anti-flagellin polyclonal antibodies (cat no. ab93713, Abcam), as previously described [13] [14] [15] [16] .
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