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Anti-GAD65抗体[GAD-6] (ab26113)

Reviews (2)Q&A (7)References (76)
Western blot - Anti-GAD65 antibody [GAD-6] (ab26113)
  • Western blot - Anti-GAD65 antibody [GAD-6] (ab26113)

Key features and details

  • Mouse monoclonal [GAD-6] to GAD65
  • Suitable for: WB
  • Reacts with: Mouse, Human
  • Isotype: IgG2a

概述

  • 产品名称

    Anti-GAD65抗体[GAD-6]
    参阅全部 GAD65 一抗

  • 描述

    小鼠单克隆抗体[GAD-6] to GAD65

  • 宿主

    Mouse

  • 特异性

    The GAD-6 monoclonal antibody ab26113 recognizes only GAD65 and has no cross reactivity with GAD67 on western blots of mouse brain or purified recombinant GAD67 and GAD65. ab26113 recognizes GAD65 primarily in nerve terminals in immunohistochemical analysis of brain sections (Kaufman et al, 1991).

  • 经测试应用

    适用于: WBmore details

  • 种属反应性

    与反应: Mouse, Human
    预测可用于: Mammals

  • 免疫原

    Full length protein (Rat)

  • 表位

    ab26113 recognizes an epitope near the C-terminus of GAD65

  • 阳性对照

    • mouse brain

  • 常规说明

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab26113于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
WB (1)
1/1000 - 1/20000. Detects a band of approximately 65 kDa (predicted molecular weight: 65 kDa).
说明
WB
1/1000 - 1/20000. Detects a band of approximately 65 kDa (predicted molecular weight: 65 kDa).

靶标

  • 功能

    Catalyzes the production of GABA.

  • 序列相似性

    Belongs to the group II decarboxylase family.

  • 翻译后修饰

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

  • 细胞定位

    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.
  • Information by UniProt

  • 数据库链接

    • 别名

      • 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

    图片

    • Western blot - Anti-GAD65 antibody [GAD-6] (ab26113)
      Western blot - Anti-GAD65 antibody [GAD-6] (ab26113)
      Anti-GAD65 antibody [GAD-6] (ab26113) at 1/5000 dilution + human brain at 15 µg

      Secondary
      rabbit anti-mouse-HRP at 1/100000 dilution

      Predicted band size: 65 kDa
      Observed band size: 65 kDa



      4-12% gradient SDS-PAGE imaged using the pico-plus ECL kit

    • Western blot - Anti-GAD65 antibody [GAD-6] (ab26113)
      Western blot - Anti-GAD65 antibody [GAD-6] (ab26113)
      Anti-GAD65 antibody [GAD-6] (ab26113) at 1/1000 dilution + mouse cerebellum at 0.75 µg

      Secondary
      HRP horse anti-mouse IgG at 1/5000 dilution

      Predicted band size: 65 kDa

    文献 (76)

    发表研究结果有使用 ab26113？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab26113 被引用在 76 文献中.

    • Hassan M  et al. Developmental Inhibitory Changes in the Primary Somatosensory Cortex of the Stargazer Mouse Model of Absence Epilepsy. Biomolecules 13:N/A (2023). PubMed: 36671571
    • Zong FJ  et al. Circadian time- and sleep-dependent modulation of cortical parvalbumin-positive inhibitory neurons. EMBO J 42:e111304 (2023). PubMed: 36477886
    • Ballon Romero SS  et al. Electroacupuncture exerts prolonged analgesic and neuroprotective effects in a persistent dental pain model induced by multiple dental pulp injuries: GABAergic interneurons-astrocytes interaction. Front Immunol 14:1213710 (2023). PubMed: 37954604
    • Tomiyama ALMR  et al. Neuroprotection by upregulation of the major histocompatibility complex class I (MHC I) in SOD1G93A mice. Front Cell Neurosci 17:1211486 (2023). PubMed: 37711512
    • Cheng YC  et al. Downregulated Calcium-Binding Protein S100A16 and HSP27 in Placenta-Derived Multipotent Cells Induce Functional Astrocyte Differentiation. Stem Cell Rev Rep 18:839-852 (2022). PubMed: 35061207
    View all Publications for this product

    客户评价及客户问答

    Show All 评价 Q&A
    提交评价 提交问题

    1-9 of 9 Abreviews or Q&A

    Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-GAD65 antibody [GAD-6]

     Average
    Abreviews
    abreview image
    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Mouse Tissue sections (brain - cortex)
    Antigen retrieval step
    None
    Permeabilization
    Yes - 0.5% Triton-X in 10% goat serum
    Specification
    brain - cortex
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    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.

    Patrick Ye

    Verified customer

    提交于 Dec 12 2016

    Western blot abreview for Anti-GAD65 antibody [GAD-6]

     Excellent
    Abreviews
    Application
    Western blot
    Sample
    Human Tissue lysate - whole (human hippocampus)
    Gel Running Conditions
    Reduced Denaturing (4-12%)
    Loading amount
    10 µg
    Treatment
    used protease inhibitors
    Specification
    human hippocampus
    Blocking step
    BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
    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.

    Abcam user community

    Verified customer

    提交于 Jun 25 2015

    Question



    Inquiry: Hello, Do you have an antigen for pre-adsorption controls for teh following product? https://www.abcam.com/GAD65-antibody-GAD-6-Neuronal-Marker-ab26113.html

    Read More
    Answer

    Thank you for the clarification! Unfortunately the blocking peptide is not available for this particular antibody. Please let me know if I can be of further assistance.

    Read More

    Question


    Thank you for replacement GAD65 ab 112497. I try it on my western blot. The ab
    lind up to all my mark. and no band was found on my positive control(Rat brain
    tissue). The reason I select 112497, because I think this is polycolonal ab and
    may give mpre chance for my cells.
    If you can send me another GAD 65 ab 75750, I can try!
    Thanks!

    Read More
    Answer

    Thank you very much for your reply.

    I am sorry to hear that the replacement ab112497 did not work in Western blot. Just to confirm, there were multiple bands in your neuron lysate, and no bands with the rab brain lysate? Were you using the same protocol as before?

    I'm sending a replacement vial of ab75750 on the order ***, which we expect to arrive around Thursday of next week. Please keep me updated about the results with this antibody as well. If you have any questions or need anything else, please let me know and I'll be happy to help.

    Read More

    Question


    Thank you for your information.
    I didn't try it in other Rat tissues, I may try it and let you know the results.
    I also couldn't find any related papers.
    I would like to try polycolone Ab.
    Best Regards!

    Read More
    Answer

    Thank you very much for your reply.

    I'm sending a free of charge vial of ab112497 on the order **** so that you can compare the results to those with ab26113. This antibody is out of stock but we expect it to arrive around September 27 (next Thursday). For our records, could you also let me know if you block with milk or BSA during your Western blot experiments?

    Please keep me updated about the results with ab112497 once you've had a chance to try it. I look forward to hearing from you. If there is anything else that we can do for you please let me know and I'll be happy to help.

    Read More

    Question

    Band around 65 kDa with rat brain tissue lysate, 40 kDa in neural cell lysate. Is this 40 kDa band specific. 1:1000 dilution. Normal culture conditions.

    Read More
    Answer

    Thank you very much for your call yesterday and for your patience while I've looked into your enquiry.

    I actually could not find a reference showing analysis of endogneous GAD65 in culture neurons by Western blot. So, I'm still not quite sure how to explain this 40 kDa band in your samples. Do you have access to any other tissues to use as a positive control, just to further verify that the antibody is binding specifically? Have you managed to find any information in the literature?

    I am very sorry that I don't have more information, but if you have any concerns about this antibody and feel that it is binding non-specifically, I'd be happy to send you a replacement antibody such as ab75750 or ab112497-

    https://www.abcam.com/GAD65-antibody-EPR2553Y-ab75750.html

    https://www.abcam.com/GAD65-antibody-ab112497.html

    I look forward to hearing from you. Please let me know if you have any questions or need anything else from me.

    Read More

    Question

    Hello,



    thanks a lot for your answer.

    I would have another question though: as the GAD65 antibody is confined to nerve terminals, is it (in general) possible to stain for GAD65 cell bodies? Are there antibodies that are specifically used to stain for these cell bodies? I mean in the literature you see stainings for GAD65 cell bodies..



    My best,

    Read More
    Answer

    Thank you for contacting us.

    We are happy to help customers find the most suitable products for their research purposes; however we are not specialized in this particular field and do not want to recommend a product that may be unsuitable. We encourage customers to consult the latest literature available through PubMed and other resources in order to find the most up-to-date information about their specific research interests.

    I am sorry that I could not be more helpful, but I hope that the available literature in this area can provide some clarification. Please do not hesitate to contact us again with other needs or with any questions about our products.

    Read More

    Question

    Lot number: Gr56581-1
    Inquiry: Hello, in our lab we want to stain for GAD65 neuronal cell bodies in mouse spinal cord (see protocol attached). As described in the literature and our protocol, we leave out any Triton steps for this. The staining works (labeling found mainly in Rexed lamina 1 and 2) but as besides the GAD65 cell bodies a lot of synapses around are labeled as well, we hardly see them! Do you know a method or a special treatment to prevent synaptic labeling in this case? For GAD67 this kind of staining works nicely (cell bodies clearly visible). Is there maybe a difference in the staining because GAD65 is supposed to be located more in synapses (in comparison to GAD67)? But as in the literature GAD65 cell body staining have been described, it should somehow be possible to make them visible in a clear way. Thanks in advance,

    Read More
    Answer

    Thank you for your enquiry regarding ab26113.

    It has been reported that GAD65 is transported to nerve terminals and this antibody (ab26113) should stain them as “puncta” around the cell body of their target neurons.

    On the other hand, GAD67 (ab26116) is localized to cell bodies, axons and terminals and it will stain these well as long as the samples undergo antigen retrieval.

    I hope this helps and if I can assist further, please do not hesitate to contact me.

    Read More

    Question

    DESCRIPTION OF THE PROBLEM No signal SAMPLE Neurodifferentiated human embryonic stem cells PRIMARY ANTIBODY Abcam (ab26113), Mouse monoclonal to GAD65, in Knouckout Serum replacement @ 1:500, incubate for 1 hour and wash 3 times with 100 uL PBS ++ DETECTION METHOD GE Healthcare In Cell 6000, High content analyzer using confocal imaging POSITIVE AND NEGATIVE CONTROLS USED As this is a routine procedure, we do not use positive and negative controls on every plate. This assay plate was stained for other markers which worked fine indicating that there was no problem with the staining method or the image capture system. We have previously used this antibody in this manner and have gotten good results. We expect our cultures to be 60-70% GAD65 positive. We have in the past run a negative control (undifferentiated stem cells). Previous run (GAD65) in red ANTIBODY STORAGE CONDITIONS stored at -20C in 10 uL aliquots FIXATION OF SAMPLE 10% formalin (Sigma-Aldrich ANTIGEN RETRIEVAL None PERMEABILIZATION STEP Yes, 0.25% triton x-100 BLOCKING CONDITIONS No blocking SECONDARY ANTIBODY Invitrogen (A31571), Donkey anti-mouse IgG Alexa Fluor 647 , in Knouckout Serum replacement @ 1:500, incubate for 1 hour and wash 3 times with 100 uL PBS ++ HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes

    Read More
    Answer

    Thank you for contacting Abcam.

    Based on the protocol the customer sent, there are a few suggestions that may help resolve the issue:

    1 - Increase the concentration of primary antibody used to 1/200.

    2 - Increase the incubation time to 3 hours at room temperature.

    I understand that the customer had previously purchased vials of this antibody and it has proved to be successful for them but as we have not tested ab26113 in either human cells or if it is not covered under our Abpromise. Please let me know if changing the above does not prove to be successful and I will if there is anything else I can do.

    Read More

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