Name: Anti-GAD65 + GAD67 antibody [EPR19366]
Brand: RabMAb
SKU: ab183999
Rating: 5 (4 reviews)

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Immunocytochemistry/ Immunofluorescence - Anti-GAD65 + GAD67 antibody [EPR19366] (ab183999)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR19366] to GAD65 + GAD67
Suitable for: IP, IHC-P, IHC-Fr, WB, ICC/IF
Reacts with: Mouse, Rat, Human

Alexa Fluor® 488 Alexa Fluor® 647 Biotin Carrier Free

製品名

Anti-GAD65 + GAD67 antibody [EPR19366]
GAD65 + GAD67 一次抗体製品一覧
製品の詳細

Rabbit monoclonal [EPR19366] to GAD65 + GAD67
由来種

Rabbit
アプリケーション

適用あり: IP, IHC-P, IHC-Fr, WB, ICC/IFmore details
種交差性

交差種: Mouse, Rat, Human
交差が予測される動物種: Common marmoset
免疫原

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
ポジティブ･コントロール
- WB: Mouse GAD67 and GAD65 fragment recombinant proteins; Human, mouse and rat cerebellum lysates; C6 whole cell lysate; Mouse and rat brain lysates. IHC-P: Mouse cerebellum and pancreas tissues; Rat cerebellum and pancreas tissues. IHC-Fr: Mouse hippocampus tissue. ICC/IF: Mouse primary neuron cells
特記事項
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.

製品の状態

Liquid
保存方法

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.
バッファー

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

Protein A purified
ポリ/モノ

モノクローナル
クローン名

EPR19366
アイソタイプ

IgG
研究分野
- Neuroscience
- Development

Alternative Versions
- Anti-GAD65 + GAD67 antibody [EPR19366] - BSA and Azide free (ab240280)
- Alexa Fluor® 488 Anti-GAD65 + GAD67 antibody [EPR19366] (ab314811)
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)

The Abpromise guarantee

Abpromise保証は、次のテスト済みアプリケーションにおけるab183999の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション	Abreviews	特記事項
IP		1/40.
IHC-P	(3)	1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr	(1)	1/2500.
WB		1/1000. Detects a band of approximately 65, 67 kDa (predicted molecular weight: 65, 67 kDa).
ICC/IF		1/100.

特記事項
IP 1/40.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr 1/2500.
WB 1/1000. Detects a band of approximately 65, 67 kDa (predicted molecular weight: 65, 67 kDa).
ICC/IF 1/100.

関連性

This gene encodes one of several forms of glutamic acid decarboxylase, identified as a major autoantigen in insulin-dependent diabetes. The enzyme encoded is responsible for catalyzing the production of gamma aminobutyric acid from L glutamic acid. A pathogenic role for this enzyme has been identified in the human pancreas since it has been identified as an autoantibody and an autoreactive T cell target in insulin dependent diabetes. This gene may also play a role in the stiff man syndrome.
細胞内局在

Cytoplasm; cytosol. Associated to cytoplasmic vesicles. In neurons, cytosolic leaflet of Golgi membranes and presynaptic clusters.
参照データベース
- Entrez Gene: 2571 Human
- Entrez Gene: 2572 Human
- Entrez Gene: 14415 Mouse
- Entrez Gene: 14417 Mouse
- Entrez Gene: 24379 Rat
- Entrez Gene: 24380 Rat
- Omim: 138275 Human
- Omim: 605363 Human
- SwissProt: Q05329 Human
- SwissProt: Q99259 Human
- SwissProt: P48318 Mouse
- SwissProt: P48320 Mouse
- SwissProt: P18088 Rat
- SwissProt: Q05683 Rat
see all
別名
- 65 kDa glutamic acid decarboxylase antibody
- 67 kDa glutamic acid decarboxylase antibody
- GAD 65 antibody
- GAD 67 antibody
- GAD1 antibody
- GAD2 antibody
- Glutamate decarboxylase 1 antibody
- Glutamate decarboxylase 2 antibody
- Glutamate decarboxylase 2 pancreatic islets and brain 65kDa antibody
- Glutamate decarboxylase 65 kDa isoform antibody
- Glutamate decarboxylase 67 kDa isoform antibody
- SCP antibody
see all

Immunocytochemistry/ Immunofluorescence - Anti-GAD65 + GAD67 antibody [EPR19366] (ab183999)

Immunocytochemistry/ Immunofluorescence analysis of mouse primary neuron cells labeling GAD65 + GAD67 with purified ab183999 at 1/100 (10 µg/mL). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab11267 Anti-MAP2 antibody [HM-2]; ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection.
Western blot - Anti-GAD65 + GAD67 antibody [EPR19366] (ab183999)

All lanes : Anti-GAD65 + GAD67 antibody [EPR19366] (ab183999) at 1/1000 dilution

Lane 1 : Mouse GAD67 fragment recombinant protein
Lane 2 : Mouse GAD65 fragment recombinant protein

Lysates/proteins at 0.01 µg per lane.

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

Predicted band size: 65, 67 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1: 1 second; Lane 2: 15 seconds.

Mouse GAD67 fragment recombinant protein contain aa468-592 with a His-Tag®. Mouse GAD65 fragment recombinant protein contain aa460-584 with a His-Tag®. These two fragment recombinant proteins were made in-house. The ~30 kDa band represents doublets of the recombinant fragments.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAD65 + GAD67 antibody [EPR19366] (ab183999)

Immunohistochemical analysis of paraffin-embedded Mouse cerebellum tissue labeling GAD65 + GAD67 with ab183999 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on mouse cerebellum is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-GAD65 + GAD67 antibody [EPR19366] (ab183999)

All lanes : Anti-GAD65 + GAD67 antibody [EPR19366] (ab183999) at 1/1000 dilution

Lane 1 : Human cerebellum lysate
Lane 2 : Mouse skin lysate
Lane 3 : Mouse lung lysate
Lane 4 : Neuro-2a (Mouse neuroblastoma cell line) whole cell lysate
Lane 5 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 6 : Mouse cerebellum lysate
Lane 7 : Rat cerebellum 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, 67 kDa
Observed band size: 65,67 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1-5: 3 minutes; Lane 6 and 7: 2 seconds.
Immunoprecipitation - Anti-GAD65 + GAD67 antibody [EPR19366] (ab183999)

Ab183999 Immunoprecipitating GAD65 + GAD67 in Human cerebellum lysate. 10µg of cell lysate was incubated with primary antibody 1/40. For western blotting Ab183999 (1/1000) was used to confirm successful immunoprecipation. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000.

Lane 1 (Input): Human cerebellum lysate 10μg
Lane 2 (+): Human cerebellum lysate with Ab183999, 1/40
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab183999 in Human cerebellum lysate
Western blot - Anti-GAD65 + GAD67 antibody [EPR19366] (ab183999)

Lane 1 : Anti-GAD65 + GAD67 antibody [EPR19366] (ab183999) at 1/5000 dilution
Lane 2 : Anti-GAD65 + GAD67 antibody [EPR19366] (ab183999) at 1/1000 dilution

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

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 65, 67 kDa
Observed band size: 65,67 kDa why is the actual band size different from the predicted?

Exposure time: 2 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAD65 + GAD67 antibody [EPR19366] (ab183999)

Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue labeling GAD65 + GAD67 with ab183999 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on mouse pancreas islets is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAD65 + GAD67 antibody [EPR19366] (ab183999)

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling GAD65 + GAD67 with ab183999 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Negative on mouse lung. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Frozen sections) - Anti-GAD65 + GAD67 antibody [EPR19366] (ab183999)

Immunohistochemical analysis of 4% paraformaldehyde fixed, 0.2% Triton X-100 permealized frozen section of Mouse hippocampus tissue labeling GAD65 + GAD67 with ab183999 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) secondary (ab150077) at 1/1000 dilution (green). The result showed mainly cytoplasmic staining on mouse hippocampus. The nuclear counterstain is DAPI (blue).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 at 1/1000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAD65 + GAD67 antibody [EPR19366] (ab183999)

Immunohistochemical analysis of paraffin-embedded Rat cerebellum tissue labeling GAD65 + GAD67 with ab183999 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on rat cerebellum is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAD65 + GAD67 antibody [EPR19366] (ab183999)

Immunohistochemical analysis of paraffin-embedded Rat pancreas tissue labeling GAD65 + GAD67 with ab183999 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on rat pancreas islet is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAD65 + GAD67 antibody [EPR19366] (ab183999)

Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling GAD65 + GAD67 with ab183999 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Negative on rat kidney. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Anti-GAD65 + GAD67 antibody [EPR19366] (ab183999)

Immunohistochemistry protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

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Datasheet download

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ab183999 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab183999 は 17 報の論文で使用されています。

Salazar JJ et al. Retinal Tissue Shows Glial Changes in a Dravet Syndrome Knock-in Mouse Model. Int J Mol Sci 24:N/A (2023). PubMed: 36769051
Wang L et al. Organizational Principles of the Centrifugal Projections to the Olfactory Bulb. Int J Mol Sci 24:N/A (2023). PubMed: 36902010
Wang M et al. Lateral septum adenosine A2A receptors control stress-induced depressive-like behaviors via signaling to the hypothalamus and habenula. Nat Commun 14:1880 (2023). PubMed: 37019936
Wong H et al. CNS-dominant human FMRP isoform rescues seizures, fear, and sleep abnormalities in Fmr1-KO mice. JCI Insight 8:N/A (2023). PubMed: 37288657
Ghuman H et al. Emergence of task-related spatiotemporal population dynamics in transplanted neurons. Nat Commun 14:7320 (2023). PubMed: 37951968

View all Publications for this product

レビューと Q&A

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レビューを送る質問を送る

1-4 of 4 Abreviews or Q&A

Application

Immunohistochemistry (Frozen sections)

Sample

Gerbil Tissue sections (Brain)

Permeabilization

Yes - 0.2% Triton for 30 minutes

Specification

Brain

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C

Fixative

Paraformaldehyde

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

投稿 Aug 17 2023

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Common marmoset Tissue sections (Brain cortex)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Citric acid

Permeabilization

Specification

Brain cortex

Blocking step

BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C

Fixative

Formaldehyde

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

投稿 Nov 26 2021

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Rat Tissue sections (Cerebellum)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Citric acid

Permeabilization

Specification

Cerebellum

Blocking step

BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C

Fixative

Formaldehyde

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

投稿 Mar 12 2021

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Mouse Tissue sections (Cerebellum)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Citric acid

Permeabilization

Specification

Cerebellum

Blocking step

BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C

Fixative

Formaldehyde

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

投稿 Mar 12 2021

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Key features and details

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製品の概要

製品名

製品の詳細

由来種

アプリケーション

種交差性

免疫原

ポジティブ･コントロール

特記事項

製品の特性

製品の状態

保存方法

バッファー

精製度

ポリ/モノ

クローン名

アイソタイプ

研究分野

関連製品

Alternative Versions

Compatible Secondaries

Isotype control

アプリケーション

The Abpromise guarantee

ターゲット情報

関連性

細胞内局在

参照データベース

別名

画像

プロトコール

データシートおよび資料

SDS download

Datasheet download

Certificate of Compliance

参考文献 (17)

レビューと Q&A

Immunohistochemistry (Frozen sections) abreview for Anti-GAD65 + GAD67 antibody [EPR19366]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-GAD65 + GAD67 antibody [EPR19366]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-GAD65 + GAD67 antibody [EPR19366]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-GAD65 + GAD67 antibody [EPR19366]