GLYT1 antibody (Glycine Transporter 1) (AA 500-600) Primary Antibody
- Target See all GLYT1 Antibodies
- AA 500-600
- Human, Mouse, Rat
- This GLYT1 antibody is un-conjugated
- Immunohistochemistry (IHC), Western Blotting (WB)
- Specific for SLC6A9.
- Whole serum
- A synthetic peptide from aa region 500-600 of human SLC6A9 conjugated to an immunogenic carrier protein was used as the antigen.
anti-Glycine Transporter 1 (GLYT1) (Middle Region) antibody Primary Antibody
GLYT1 Reactivity: Human, Mouse, Rat, Cow, Dog, Guinea Pig, Horse, Rabbit, Zebrafish (Danio rerio) WB Host: Rabbit Polyclonal unconjugated
- Application Notes
A dilution of 1: 300 to 1: 2000 is recommended.
The optimal dilution should be determined by the end user.
Not yet tested in other applications.
- For Research Use only
Validation #100063 (Immunohistochemistry)Validation ImagesA cross section of a mouse retina labelled with ABIN1841935 (rabbit polyclonal anti-glycine transporter 1, GLYT-1 1:400) and a Alexa488 conjugated secondary donkey anti-rabbit antibody 1:400(green). ABIN1841935 specifically labels glycinergic amacrine cells in the mouse retina with no unspecific labelling in other retinal layers. Nuclei are labelled with DAPI (blue).Full Methods
- Kleinlogel lab, Institute of Physiology, University of Bern, Switzerland
- Glycine Transporter 1 (GLYT1) (AA 500-600)
- Lot Number
- Method validated
- Positive Control
- Glycinergic amacrine cells in the inner nuclear layer and the inner plexiform layer of the mouse retina.
- Negative Control
- Other ocular tissue in the same sections.
- This antibody worked very well in the mouse retina. ABIN1841935 specifically labels glycinergic amacrine cells – which have a characteristic morphology - in the mouse retina with no unspecific labelling in other retinal layers.
- Primary Antibody
- Secondary Antibody
- Life Technologies, A11008, lot 1761258
- Full Protocol
- Immediately nucleate eyes from euthanized mice.
- Hemi-sect the eyes around the border of the cornea and remove the lens.
- Wash the eye cups in 1x PBS.
- Fix tissue in 4% PFA for 30-40min at RT.
- Thoroughly wash PFA from tissue twice in 1x PBS for 5 min at RT.
- Cryoprotection with Sucrose:
- Immerse tissue in 10% Sucrose (in 1x PBS) for 2h at RT.
- Immerse tissue in 20% Sucrose (in 1x PBS) for 2h at RT.
- Immerse tissue in 30% Sucrose (in 1x PBS) overnight at 4°C.
- Fill cryomold with OCT compound (Sakura Tissue Tek, 4583 lot 2317).
- Put 2-Methylbutan into a big glass beaker, and beaker into liquid nitrogen.
- Wash tissue briefly in 1x PBS after cryoprotection.
- Place tissue into OCT compound in cryomold “horizontally” and at middle depth.
- Place cryomold into cold 2-methylbutane; take care that it is not submerged, otherwise the OCT block may crack.
- After approximately 30sec-1 min, take cryomold out and store it at -20°C for sectioning.
- Slide Preparation:
- Let the frozen slides thaw and dry at RT 10-30min.
- Remove OCT by washing slides twice in a 1x PBS baths for some minutes.
- Dry around sections with paper tissue then draw a big enough circle with a wax pen on the slide around the sections.
- Cover the sections with 0.1 M Gly/PBS for 30min to block unspecific aldehyde groups.
- Remove Gly/PBS and wash three times for at least 5min with 1x PBS.
- Block sections with 2x normal goat serum (6% NGS, 2% BSA, 1% TritonX-100) for 30min.
- Primary AB:
- Dilute primary rabbit anti glycine transporter 1 (GLYT1) antibody (ABIN1841935, lot Rb1929-281012-WS) in 1x NGS (see above) to 1:400. Centrifuge the solution at 16.000rcf for 6min to pellet aggregates.
- Add 30µL of the primary antibody solution from the top of the supernatant on each section, wet paper towels in the incubation chamber.
- Insert the slides into the incubation chamber. Prevent the sections from drying out!
- Incubate overnight at 4°C.
- Secondary AB:
- Wash slides four times for at least 5min in 1x PBS.
- Dilute secondary goat anti rabbit AF488 conjugated antibody (Life Technologies, A11008, lot 1761258) in 1x NGS to 1:400. Centrifuge the solution at 16.000rcf for 6min to pellet aggregates.
- Add 1:2000 DAPI (from a 1mg/ml stock dilution) to the solution.
- Add 30µL of the secondary antibody solution from the top of the supernatant on each section.
- Uncubate for 2h at RT in the dark.
- Wash and mount:
- Wash slides four times for at least 5min in 1x PBS.
- Add a small bubble-free drop of fluorescence mounting medium (DAKO, S3023, lot 10115314) on top of the wet sections.
- Place a coverslip on it avoiding bubbles. Seal the edges of the coverslip with a clear nail polish.
- Incubate over night at 4°C and in the dark.
- Image acquisition on a Zeiss 880 laser scanning microscope using automated filter settings for AF488, a 40x oil immersion objective, a laser intensity of 10% and a gain of approximately 400.
- Experimental Notes
- Reconstitute in 100 µL of sterile water. Centrifuge to remove any insoluble material.
- Handling Advice
- Avoid freeze and thaw cycles.
- 4 °C/-20 °C
- Storage Comment
- Maintain the lyophilised/reconstituted antibodies frozen at -20°C for long term storage and refrigerated at 2-8°C for a shorter term. When reconstituting, glycerol (1:1) may be added for an additional stability. Avoid freeze and thaw cycles.
- Expiry Date
- 12 months
van Wyk, Hulliger, Girod, Ebneter, Kleinlogel: "Present Molecular Limitations of ON-Bipolar Cell Targeted Gene Therapy." in: Frontiers in neuroscience, Vol. 11, pp. 161, 2017 (PubMed).
- van Wyk, Hulliger, Girod, Ebneter, Kleinlogel: "Present Molecular Limitations of ON-Bipolar Cell Targeted Gene Therapy." in: Frontiers in neuroscience, Vol. 11, pp. 161, 2017 (PubMed).
- Alternative Name
- SLC6A9 (GLYT1 Products)
- GLYT1, Glyt-1, Glyt1, GLYT-1, GLYT-1b, glyt1, slc6a9, glyT-1, glyT1, solute carrier family 6 member 9, solute carrier family 6 (neurotransmitter transporter, glycine), member 9, glycine transporter 1, SLC6A9, Slc6a9, glyt1, slc6a9
FUNCTION: Terminates the action of glycine by its high affinity sodium-dependent reuptake into presynaptic terminals. May play a role in regulation of glycine levels in NMDA receptor-mediated neurotransmission. LOCATION: Membrane, Multi-pass membrane protein. Tissue specificity: At E11, expressed in the ventral part of the ventricular zone. At E15, also expressed in adjacent mantle tissue and the meninges. Strongly expressed in E12 and E15 liver.
Synonyms: Sodium- and chloride-dependent glycine transporter 1, GlyT-1, GlyT1, Solute carrier family 6 member 9