Glycine Transporter 1 (GLYT1) (AA 500-600) antibody

Details for Product No. ABIN1841935, Supplier: Log in to see
Antigen
  • glyt1
  • slc6a9
  • GLYT-1
  • GLYT-1b
  • Glyt1
  • GLYT1
  • Glyt-1
  • glyT-1
  • glyT1
  • glycine transporter 1
  • solute carrier family 6 (neurotransmitter transporter, glycine), member 9
  • glyt1
  • Slc6a9
  • SLC6A9
Epitope
AA 500-600
9
3
2
1
1
Reactivity
Human, Mouse (Murine), Rat (Rattus)
25
24
22
5
5
4
4
4
1
1
1
Host
Rabbit
30
1
Clonality
Polyclonal
Conjugate
Un-conjugated
1
1
1
1
1
1
1
1
1
1
1
1
1
1
Application
Immunohistochemistry (IHC), Western Blotting (WB)
17
13
5
5
1
Options
Supplier
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Available images

'Independent Validation' Badge
Antigen Glycine Transporter 1 (GLYT1) (AA 500-600)
Lot Number Rb1929-281012-WS
Method validated Immunohistochemistry
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.
Notes 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 ABIN1841935
Secondary Antibody Life Technologies, A11008, lot 1761258
Protocol
  • Fixation:
    • 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.
  • Freezing:
    • 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.
  • Microscopy:
    • 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.
Validation Images
Immunohistochemistry validation image for anti-Glycine Transporter 1 (GLYT1) (AA 500-600) antibody (ABIN1841935) A cross section of a mouse retina labelled with ABIN1841935 (rabbit polyclonal anti-g...
Immunogen A synthetic peptide from aa region 500-600 of human SLC6A9 conjugated to an immunogenic carrier protein was used as the antigen.
Specificity Specific for SLC6A9.
Purification Whole serum
Alternative Name SLC6A9 (GLYT1 Antibody Abstract)
Background 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
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.
Restrictions For Research Use only
Format Lyophilized
Reconstitution Reconstitute in 100 µL of sterile water. Centrifuge to remove any insoluble material.
Handling Advice Avoid freeze and thaw cycles.
Storage 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