[Preparation and application of rabbit polyclonal antibody against human glutamine fructose-6-phosphate amidotransferase 1(GFPT1)]. 2022

Shaochun Wu, and Lin Liu, and Yanchun Gao, and Yuchao Gu
School of Medicine and Pharmacy, Ocean University of China, Qingdao 266000, China.

Objective To prepare rabbit polyclonal antibody specifically against human glutamine fructose-6-phosphate amidotransferase 1(GFPT1). Methods The protein sequences of GFPT1 and its highly homologous isozyme GFPT2 were compared. Two peptides for the specific sequence of GFPT1 were designed and synthesized. New Zealand rabbits were immunized by peptide coupled with Keyhole Limpet hemocyanin (KLH) as antigen. Antiserum was obtained after 3 booster immunizations. The titer of the antiserum against GFPT1 were detected by ELISA. The E.coli expression vectors of GFPT1 and GFPT2 were constructed, and the recombinant proteins of GFPT1 and GFPT2 were obtained by induced expression. GFPT1 and GFPT2 recombinant proteins were analyzed by Western blot to verify the specificity of the antiserum. Immuno-fluorescence cytochemical staining for GFPT1 expression in 786-O cells was verified as for whether the obtained antiserum could recognize the endogenously expressed GFPT1. Results Polyclonal antibody specifically recognizing GFPT1 was obtained and the titer of polyclonal antibody reached 1:1 458 000. Conclusion The experiment successfully prepared the specific rabbit polyclonal antibody against GFPT1.

UI MeSH Term Description Entries
D007106 Immune Sera Serum that contains antibodies. It is obtained from an animal that has been immunized either by ANTIGEN injection or infection with microorganisms containing the antigen. Antisera,Immune Serums,Sera, Immune,Serums, Immune
D011817 Rabbits A burrowing plant-eating mammal with hind limbs that are longer than its fore limbs. It belongs to the family Leporidae of the order Lagomorpha, and in contrast to hares, possesses 22 instead of 24 pairs of chromosomes. Belgian Hare,New Zealand Rabbit,New Zealand Rabbits,New Zealand White Rabbit,Rabbit,Rabbit, Domestic,Chinchilla Rabbits,NZW Rabbits,New Zealand White Rabbits,Oryctolagus cuniculus,Chinchilla Rabbit,Domestic Rabbit,Domestic Rabbits,Hare, Belgian,NZW Rabbit,Rabbit, Chinchilla,Rabbit, NZW,Rabbit, New Zealand,Rabbits, Chinchilla,Rabbits, Domestic,Rabbits, NZW,Rabbits, New Zealand,Zealand Rabbit, New,Zealand Rabbits, New,cuniculus, Oryctolagus
D011994 Recombinant Proteins Proteins prepared by recombinant DNA technology. Biosynthetic Protein,Biosynthetic Proteins,DNA Recombinant Proteins,Recombinant Protein,Proteins, Biosynthetic,Proteins, Recombinant DNA,DNA Proteins, Recombinant,Protein, Biosynthetic,Protein, Recombinant,Proteins, DNA Recombinant,Proteins, Recombinant,Recombinant DNA Proteins,Recombinant Proteins, DNA
D004926 Escherichia coli A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc. Alkalescens-Dispar Group,Bacillus coli,Bacterium coli,Bacterium coli commune,Diffusely Adherent Escherichia coli,E coli,EAggEC,Enteroaggregative Escherichia coli,Enterococcus coli,Diffusely Adherent E. coli,Enteroaggregative E. coli,Enteroinvasive E. coli,Enteroinvasive Escherichia coli
D005636 Fructosephosphates
D005945 Glutamine-Fructose-6-Phosphate Transaminase (Isomerizing) An enzyme that catalyzes the synthesis of fructose-6-phosphate plus GLUTAMINE from GLUTAMATE plus glucosamine-6-phosphate. Glucosamine Synthetase,Glucosaminephosphate Isomerase (Glutamine-Forming),Hexosephosphate Aminotransferase,2-Amino-2-Deoxy-D-Glucose-6-Phosphate Ketol-Isomerase,Glucosamine 6-Phosphate Synthetase,Glucosamine-6-Phosphate Synthase,Glucosaminephosphate Isomerase (Glutamine Forming),Glutamine-Fructose-6-P Aminotransferase,Glutamine-Fructose-6-Phosphate Aminotransferase,Glutamine:Fructose-6-Phosphate-Amidotransferase,2 Amino 2 Deoxy D Glucose 6 Phosphate Ketol Isomerase,6-Phosphate Synthetase, Glucosamine,Aminotransferase, Glutamine-Fructose-6-P,Aminotransferase, Glutamine-Fructose-6-Phosphate,Aminotransferase, Hexosephosphate,Glucosamine 6 Phosphate Synthase,Glucosamine 6 Phosphate Synthetase,Glutamine Fructose 6 P Aminotransferase,Glutamine Fructose 6 Phosphate Aminotransferase,Glutamine:Fructose 6 Phosphate Amidotransferase,Ketol-Isomerase, 2-Amino-2-Deoxy-D-Glucose-6-Phosphate
D005973 Glutamine A non-essential amino acid present abundantly throughout the body and is involved in many metabolic processes. It is synthesized from GLUTAMIC ACID and AMMONIA. It is the principal carrier of NITROGEN in the body and is an important energy source for many cells. D-Glutamine,L-Glutamine,D Glutamine,L Glutamine
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D000906 Antibodies Immunoglobulin molecules having a specific amino acid sequence by virtue of which they interact only with the ANTIGEN (or a very similar shape) that induced their synthesis in cells of the lymphoid series (especially PLASMA CELLS).

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