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

GLYT1 Reactivity: Human, Mouse, Rat IHC, WB Host: Rabbit Polyclonal unconjugated
Pubmed (1) Independent Validation (1)
Catalog No. ABIN1841935
$454.67
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100 μL
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  • Target See all GLYT1 Antibodies
    GLYT1
    Binding Specificity
    • 15
    • 5
    • 2
    • 1
    • 1
    • 1
    • 1
    AA 500-600
    Reactivity
    • 25
    • 24
    • 23
    • 5
    • 5
    • 5
    • 5
    • 4
    • 2
    • 1
    • 1
    Human, Mouse, Rat
    Host
    • 25
    • 1
    Rabbit
    Clonality
    • 26
    Polyclonal
    Conjugate
    • 10
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    This GLYT1 antibody is un-conjugated
    Application
    • 14
    • 13
    • 13
    • 7
    • 3
    • 3
    • 1
    • 1
    • 1
    Immunohistochemistry (IHC), Western Blotting (WB)
    Specificity
    Specific for SLC6A9.
    Purification
    Whole serum
    Immunogen
    A synthetic peptide from aa region 500-600 of human SLC6A9 conjugated to an immunogenic carrier protein was used as the antigen.
  • 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
  • Validation #100063 (Immunohistochemistry)
    'Independent Validation' Badge
    by
    Kleinlogel lab, Institute of Physiology, University of Bern, Switzerland
    No.
    #100063
    Date
    11/20/2016
    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.
    'Independent Validation' Badge
    Validation Images
    Full Methods
    Primary Antibody
    ABIN1841935
    Secondary Antibody
    Life Technologies, A11008, lot 1761258
    Full 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.
    Experimental Notes
  • 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
  • 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).

  • Target
    GLYT1
    Alternative Name
    SLC6A9 (GLYT1 Products)
    Synonyms
    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
    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
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