Overview

  • Product name

    Anti-VGluT1 antibody [EPR22269]
    See all VGluT1 primary antibodies
  • Description

    Rabbit monoclonal [EPR22269] to VGluT1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, IHC-Fr, IPmore details
    Unsuitable for: ICC/IF
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Mouse VGluT1 aa 500 to the C-terminus. The exact sequence is proprietary.
    Database link: Q3TXX4

  • Positive control

    • WB: Human, mouse and rat brain lysates. IHC-P: Mouse cerebrum tissue; Rat and human cerebral cortex tissues. IHC-Fr: Mouse and rat hippocampus tissues. IP: Mouse brain lysate.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab227805 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000. Detects a band of approximately 62 kDa (predicted molecular weight: 62 kDa).
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr 1/100.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)

IP 1/30.
  • Application notes
    Is unsuitable for ICC/IF.
  • Target

    • Function

      Mediates the uptake of glutamate into synaptic vesicles at presynaptic nerve terminals of excitatory neural cells. May also mediate the transport of inorganic phosphate.
    • Tissue specificity

      Expressed in several regions of the brain including amygdala, cerebellum, cerebral cortex, hippocampus, frontal lobe, medulla, occipital lobe, putamen and temporal lobe.
    • Sequence similarities

      Belongs to the major facilitator superfamily. Sodium/anion cotransporter family. VGLUT subfamily.
    • Cellular localization

      Cytoplasmic vesicle, secretory vesicle, synaptic vesicle membrane. Membrane. Cell junction, synapse, synaptosome.
    • Information by UniProt
    • Database links

    • Alternative names

      • BNPI antibody
      • Brain specific Na (+) dependent inorganic phosphate cotransporter antibody
      • Brain specific Na dependent inorganic phosphate cotransporter antibody
      • Brain-specific Na(+)-dependent inorganic phosphate cotransporter antibody
      • Slc17a7 antibody
      • Solute carrier family 17 (sodium-dependent inorganic phosphate cotransporter), member 7 antibody
      • Solute carrier family 17 member 7 antibody
      • Vesicular glutamate transporter 1 antibody
      • VGLU1_HUMAN antibody
      • VGLUT 1 antibody
      • VGluT1 antibody
      see all

    Images

    • All lanes : Anti-VGluT1 antibody [EPR22269] (ab227805) at 1/1000 dilution

      All lanes : Mouse brain lysate

      Lysates/proteins at 20 µg/ml per lane.

      Secondary
      Lane 1 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
      Lanes 2-3 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

      Predicted band size: 62 kDa
      Observed band size: 62 kDa


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

      Different lysate preparation and boiling methods resulted in different banding patterns observed. The lysates in lanes 1 and 3 were prepared by RIPA lysis method with and without boiling. The lysate in lane 2 was prepared by 1% SDS Hot lysis method. For WB sample preparation, we recommend using RIPA lysis buffer and without boiling.

       

    • Immunohistochemical analysis of paraffin-embedded human cerebral cortex tissue labeling VGluT1 with ab227805 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on human cerebral cortex (PMID: 29532891). Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemical analysis of paraffin-embedded rat cerebral cortex tissue labeling VGluT1 with ab227805 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on rat cerebral cortex (PMID: 29532891) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling VGluT1 with ab227805 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Specific cytoplasmic staining on mouse hippocampus, positive staining was also observed on mouse cerebral cortex and thalamus (PMID: 29532891). Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen rat hippocampus tissue labeling VGluT1 with ab227805  at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Cytoplasmic staining on rat hippocampus (PMID: 29532891) is observed.

      The nuclear counter stain is DAPI (blue).

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse hippocampus tissue labeling VGluT1 with ab227805 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Cytoplasmic staining on mouse hippocampus (PMID: 29532891) is observed.

      The nuclear counter stain is DAPI (blue).

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • VGluT1 was immunoprecipitated from 0.35 mg of mouse brain lysate with ab227805 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab227805 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

      Lane 1: Mouse brain lysate 10 µg (Input). 

      Lane 2: ab227805 IP in mouse brain lysate. 

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab227805 in mouse brain lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 1 second.

      The pattern of oligomeric/dimeric forms observed is consistent with what has been described in the literature (PMID: 15192755).

    • Anti-VGluT1 antibody [EPR22269] (ab227805) at 1/1000 dilution + Human brain lysate at 20 µg

      Secondary
      VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

      Predicted band size: 62 kDa
      Observed band size: 62 kDa


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

      Oligomeric forms observed in the human brain lysate were due to the 1% SDS Hot lysate preparation method, consistent with what has been described in the literature (PMID: 15192755).

    • All lanes : Anti-VGluT1 antibody [EPR22269] (ab227805) at 1/1000 dilution

      Lane 1 : Mouse brain lysate
      Lane 2 : Rat brain lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Lane 1 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
      Lane 2 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

      Predicted band size: 62 kDa
      Observed band size: 62 kDa


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

      Brain lysates were prepared with RIPA lysis buffer. The pattern of oligomers/dimers observed is consistent with what has been described in the literature (PMID: 15192755).

    • Immunohistochemical analysis of paraffin-embedded human liver tissue labeling VGluT1 with ab227805 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Counter stained with hematoxylin.

      Negative control: No staining on human liver is observed.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling VGluT1 with ab227805 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Counter stained with hematoxylin.

      Negative control: No staining on rat liver is observed.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling VGluT1 with ab227805 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Counter stained with hematoxylin.

      Negative control: No staining on mouse liver.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    References

    ab227805 has not yet been referenced specifically in any publications.

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