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Guinea pig anti glyt2

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Guinea pig anti-GlyT2 is a laboratory reagent used for the detection and quantification of the glycine transporter 2 (GlyT2) protein in biological samples. It is a specific antibody raised in guinea pigs against the GlyT2 protein, which is involved in the regulation of glycine neurotransmission in the central nervous system.

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Lab products found in correlation

Guinea pig anti vglut1, by Merck Group (1 mentions) Dako antibody diluent, by Agilent Technologies (1 mentions) Alexa fluor 488 affinipure donkey anti guinea pig igg, by Jackson ImmunoResearch (1 mentions) Alexa fluor 594 affinipure donkey anti rabbit igg, by Jackson ImmunoResearch (1 mentions) Anti fading medium, by Agilent Technologies (1 mentions)

2 protocols using guinea pig anti glyt2

1

Antibody Production and Characterization

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The mouse Crygn peptide CDNFQDQGFMNRVN that was predicted to be antigenic and to discriminate among related proteins was synthesized at Pineda (Berlin, Germany), conjugated to carrier, and used to immunize rabbits. The rabbit anti-Crygd/e was produced by Genescript (Piscataway Township, NJ, USA) by immunization with the peptide RGFQGRHYECSTDHC that is present in both Crygd and Cryg/e. Antibodies were cleaned up by affinity purification. The primary antibody rabbit anti-Cryg was kindly provided by Dr. Samuel Zigler. All three primary antibodies were used in the following dilutions: immunohistochemistry: 1:50 for anti-Crygd/e and Crygn; 1:200 for anti-Cryg, immunocytochemistry: 1:100, and immunoblotting 1:250 for all three antibodies. The primary antibodies guinea pig anti-GlyT2 (Millipore; Darmstadt, Germany) (1:1,000), guinea pig anti-VGluT1 (Millipore) (1:5,000) and the rabbit anti-GlyT2 (1:1,000) have been characterized previously [59 (link),60 (link)]. Mouse anti-flag was obtained from Genescript (1:500).
Hartwich H., Rosengauer E., Rüttiger L., Wilms V., Waterholter S.K, & Nothwang H.G. (2016). Functional Role of γ-Crystallin N in the Auditory Hindbrain. PLoS ONE, 11(8), e0161140.
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2

Dual Immunostaining of c-Fos and GlyT2

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Every fourth section of tissue was used for immunofluorescence double-staining for c-Fos and GlyT2 in each brain. Brain sections were blocked in 5% normal donkey serum in PBS- Triton X-100 (PBS-T) for 1h, and then incubated in a mixed solution of guinea pig anti-GlyT2 (1:1000 dilution, Millipore; Billerica, MA) and rabbit anti-c-Fos primary antibodies (1:200 dilution, Santa Cruz Biotechnology; Santa Cruz, CA) diluted in DAKO antibody diluents (DAKO; Carpinteria, CA) for 36 hrs. After incubation with primary antibodies, PBS-T was used to wash brain sections. After three times of wash, brain tissue was incubated in secondary antibodies (1:200 dilution for Alexa Fluor® 488-AffiniPure Donkey Anti-Guinea Pig IgG and 1:100 dilution for Alexa Fluor 594-AffiniPure Donkey Anti-Rabbit IgG; Jackson ImmunoResearch; West Grove, PA) for 2 hrs. Slices of tissue were washed with PBS-T three times and then mounted on glass slides. Sides were air-dried and cover-slipped with anti-fading medium (DAKO; Carpinteria, CA). Negative controls were performed in the absence of the primary antibody and the results showed no c-Fos or GlyT2 positive staining.
Tsai H.W., Condrey J., Adams S, & Davenport P.W. (2014). THE EFFECT OF TRACHEAL OCCLUSION ON RESPIRATORY LOAD COMPENSATION: CHANGES IN NEURONS CONTAINING INHIBITORY NEUROTRANSMITTER IN THE NUCLEUS OF THE SOLITARY TRACT IN CONSCIOUS RATS. Respiratory physiology & neurobiology, 204, 138-146.
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