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  1. Link

    gad65-antibody-epr22952-70-ab239372.pdf

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Neuroscience Neurotransmitter Amino Acids Glutamate
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RecombinantRabMAb

Recombinant Anti-GAD65 antibody [EPR22952-70] (ab239372)

  • Datasheet
  • SDS
  • Certificate of Compliance
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Western blot - Anti-GAD65 antibody [EPR22952-70] (ab239372)
  • Immunohistochemistry (Frozen sections) - Anti-GAD65 antibody [EPR22952-70] (ab239372)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAD65 antibody [EPR22952-70] (ab239372)
  • Immunohistochemistry (Frozen sections) - Anti-GAD65 antibody [EPR22952-70] (ab239372)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAD65 antibody [EPR22952-70] (ab239372)
  • Immunocytochemistry/ Immunofluorescence - Anti-GAD65 antibody [EPR22952-70] (ab239372)
  • Flow Cytometry (Intracellular) - Anti-GAD65 antibody [EPR22952-70] (ab239372)
  • Immunoprecipitation - Anti-GAD65 antibody [EPR22952-70] (ab239372)
  • Anti-GAD65 antibody [EPR22952-70] (ab239372)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22952-70] to GAD65
  • Suitable for: Flow Cyt (Intra), IP, IHC-P, WB, IHC-Fr, ICC/IF
  • Reacts with: Mouse, Rat

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Carrier Free

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Overview

  • Product name

    Anti-GAD65 antibody [EPR22952-70]
    See all GAD65 primary antibodies
  • Description

    Rabbit monoclonal [EPR22952-70] to GAD65
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), IP, IHC-P, WB, IHC-Fr, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Mouse brain and cerebellum tissue lysates. Rat brain and cerebellum tissue lysates IHC-P: Rat pancreas and Mouse cerebrum tissues. IHC-Fr: Rat pancreas and Mouse cerebrum tissues. Flow Cyt (intra): Beta-TC-6IP: Mouse brain lysate
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR22952-70
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurotransmitter
    • Amino Acids
    • Glutamate
    • Neuroscience
    • Neurology process
    • Metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Amino acid metabolism
    • Neuroscience
    • Development

Associated products

  • Alternative Versions

    • Anti-GAD65 antibody [EPR22952-70] - BSA and Azide free (ab270037)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Related Products

    • Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)
    • Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab239372 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
Flow Cyt (Intra)
1/60.
IP
1/30.
IHC-P (2)
1/2000.
WB
1/5000. Predicted molecular weight: 65 kDa.
IHC-Fr
1/100.
ICC/IF
1/50.
Notes
Flow Cyt (Intra)
1/60.
IP
1/30.
IHC-P
1/2000.
WB
1/5000. Predicted molecular weight: 65 kDa.
IHC-Fr
1/100.
ICC/IF
1/50.

Target

  • Function

    Catalyzes the production of GABA.
  • Sequence similarities

    Belongs to the group II decarboxylase family.
  • Post-translational
    modifications

    Phosphorylated; which does not affect kinetic parameters or subcellular location.
    Palmitoylated; which is required for presynaptic clustering.
  • Cellular localization

    Cytoplasm > cytosol. Cytoplasmic vesicle. Cell junction > synapse > presynaptic cell membrane. Golgi apparatus membrane. Associated to cytoplasmic vesicles. In neurons, cytosolic leaflet of Golgi membranes and presynaptic clusters.
  • Target information above from: UniProt accession Q05329 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 14417 Mouse
    • Entrez Gene: 24380 Rat
    • SwissProt: P48320 Mouse
    • SwissProt: Q05683 Rat
    • Unigene: 4784 Mouse
    • Unigene: 29951 Rat
    • Alternative names

      • 65 kDa glutamic acid decarboxylase antibody
      • DCE 2 antibody
      • DCE2 antibody
      • DCE2_HUMAN antibody
      • GAD 2 antibody
      • GAD 65 antibody
      • GAD-2 antibody
      • GAD-65 antibody
      • Gad2 antibody
      • Glutamate decarboxylase 2 (pancreas) antibody
      • Glutamate Decarboxylase 2 (pancreatic islets and brain 65kDa) antibody
      • Glutamate decarboxylase 2 antibody
      • Glutamate Decarboxylase 65 antibody
      • Glutamate decarboxylase 65 kDa isoform antibody
      • Glutamic Acid Decarboxylase 2 antibody
      • Glutamic Acid Decarboxylase 65 antibody
      • MGC161605 antibody
      • MGC161607 antibody
      see all

    Images

    • Western blot - Anti-GAD65 antibody [EPR22952-70] (ab239372)
      Western blot - Anti-GAD65 antibody [EPR22952-70] (ab239372)
      All lanes : Anti-GAD65 antibody [EPR22952-70] (ab239372) at 1/5000 dilution

      Lane 1 : Mouse brain tissue lysate
      Lane 2 : Mouse cerebellum tissue lysate
      Lane 3 : Rat brain tissue lysate
      Lane 4 : Rat cerebellum tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 65 kDa



      Blocking/diluting buffer and concentration: 5% NFDM/TBST

      Exposure time: 8 seconds.

      Bands above 100kDa are multimers of GAD65.

      The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 10601283, 21734072).

    • Immunohistochemistry (Frozen sections) - Anti-GAD65 antibody [EPR22952-70] (ab239372)
      Immunohistochemistry (Frozen sections) - Anti-GAD65 antibody [EPR22952-70] (ab239372)

      Immunohistochemical analysis of 4% PFA fixed 0.2% Triton X-100 permeabilized frozen Rat pancreas tissue labeling GAD65 with ab239372 at 1/100 dilution (Green) followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution. The nuclear counterstain was DAPI (Blue).

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody was ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAD65 antibody [EPR22952-70] (ab239372)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAD65 antibody [EPR22952-70] (ab239372)

      Immunohistochemical analysis of paraffin-embedded Rat pancreas tissue labeling GAD65 with ab239372 at 1/2000 (0.323 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on islet of rat pancreas. The section was incubated with ab239372 for 15 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    • Immunohistochemistry (Frozen sections) - Anti-GAD65 antibody [EPR22952-70] (ab239372)
      Immunohistochemistry (Frozen sections) - Anti-GAD65 antibody [EPR22952-70] (ab239372)

      Immunohistochemical analysis of 4% PFA fixed 0.2% Triton X-100 permeabilized frozen Mouse cerebellum tissue labeling GAD65 with ab239372 at 1/100 dilution (Green) followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution. The nuclear counterstain was DAPI (Blue). Positive staining on mouse cerebellum is observed.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody was ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAD65 antibody [EPR22952-70] (ab239372)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAD65 antibody [EPR22952-70] (ab239372)

      Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling GAD65 with ab239372 at 1/2000 (0.323 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse cerebrum is observed. The section was incubated with ab239372 for 15 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    • Immunocytochemistry/ Immunofluorescence - Anti-GAD65 antibody [EPR22952-70] (ab239372)
      Immunocytochemistry/ Immunofluorescence - Anti-GAD65 antibody [EPR22952-70] (ab239372)

      Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron cells labelling GAD65 with ab239372 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in mouse primary neurons.is observed. ab195889 Anti-alpha Tubulin antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/1000 dilution (Red). The Nuclear counterstain was DAPI (Blue). Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection.

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

    • Flow Cytometry (Intracellular) - Anti-GAD65 antibody [EPR22952-70] (ab239372)
      Flow Cytometry (Intracellular) - Anti-GAD65 antibody [EPR22952-70] (ab239372)

      Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Beta-TC-6 (Mouse pancreas insulinoma beta cell) cells labelling GAD65 with ab239372 at 1/60 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. 

       

       

    • Immunoprecipitation - Anti-GAD65 antibody [EPR22952-70] (ab239372)
      Immunoprecipitation - Anti-GAD65 antibody [EPR22952-70] (ab239372)

      GAD65 was immunoprecipitated from 0.35 mg Mouse brain lysate with ab239372 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab239372 at 1/1000 dilution (0.45 μg/ml). VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

      Lane 1: Mouse brain lysate 10 μg

      Lane 2: ab239372 IP in Mouse brain lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab239372 in Mouse brain lysate.

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

      Exposure time: 3 seconds

      Bands above 100kDa are multimers of GAD65 (PMID: 10601283).

    • Anti-GAD65 antibody [EPR22952-70] (ab239372)
      Anti-GAD65 antibody [EPR22952-70] (ab239372)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (0)

    Publishing research using ab239372? Please let us know so that we can cite the reference in this datasheet.

    ab239372 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-2 of 2 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-GAD65 antibody [EPR22952-70]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (Cerebellum)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citric acid
    Permeabilization
    No
    Specification
    Cerebellum
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    MR. Carl Hobbs

    Verified customer

    Submitted Mar 03 2020

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-GAD65 antibody [EPR22952-70]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Cerebellum)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citric acid
    Permeabilization
    No
    Specification
    Cerebellum
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    MR. Carl Hobbs

    Verified customer

    Submitted Mar 03 2020

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