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product name
Anti-Glutamate Decarboxylase Antibody, 65 kDa isoform, clone GAD-6, clone GAD-6, Chemicon®, from mouse
由来生物
mouse
品質水準
抗体製品の状態
purified immunoglobulin
抗体製品タイプ
primary antibodies
クローン
GAD-6, monoclonal
化学種の反応性
human, rat
メーカー／製品名
Chemicon®
テクニック
immunohistochemistry: suitable
western blot: suitable
アイソタイプ
IgG2a
NCBIアクセッション番号
UniProtアクセッション番号
輸送温度
dry ice
ターゲットの翻訳後修飾
unmodified
遺伝子情報
human ... GAD2(2572)
詳細
Gutamic acid decarboxylase (GAD; E.C. 4.1.1.15) is the enzyme responsible for the conversion of glutamic acid to gamma-aminobutyric acid (GABA), the major inhibitory transmitter in higher brain regions, and putative paracrine hormone in pancreatic islets. Two molecular forms of GAD (65kDa and 67kDa, 64% aa identity between forms) are highly conserved and both forms are expressed in the CNS, pancreatic islet cells, testis, oviduct and ovary. The isoforms are regionally distributed cytoplasmically in the brains of rats and mice (Sheikh, S. et al. 1999). GAD65 is an ampiphilic, membrane-anchored protein (585aa), encoded on human chromosome 10, and is responsible for vesicular GABA production. GAD67 is cytoplasmic (594aa.), encoded on chromosome 2, and seems to be responsible for significant cytoplasmic GABA production. GAD expression changes during neural development in rat spinal cord. GAD65 is expressed transiently in commissural axons around E13 but is down regulated the next day while GAD67 expression increases mostly in the somata of those neurons (Phelps, P. et al. 1999). In mature rat pancreas, GAD65 and GAD67 appear to be differentially localized, GAD65 primarily in insulin-containing beta cells and GAD67 in glucagon-containing (A) cells (Li, L. et al. 1995). GAD67 expression seems to be particularly plastic and can change in response to experimental manipulation (for example neuronal stimulation or transection) or disease progression and emergent disorders like schizophrenia (Volk et al., 2000). Colocalization of the two GAD isoforms also shows changes in GAD65/GAD67 distributions correlated with certain disease states such as IDDM and SMS.
特異性
Recognizes the lower molecular weight isoform of the two GAD isoforms identified in brain (Gottlieb, et al., 1986; Chang & Gottlieb, 1988). This monclonal antibody can be used for immunohistochemical localization in brain or pancreas. Anti-GAD has also been used to label purified GAD on Western blots (Chang & Gottlieb, 1988).
免疫原
Purified rat brain GAD.
アプリケーション
Research Category
ニューロサイエンス
ニューロサイエンス
Research Sub Category
神経伝達物質及び受容体
神経伝達物質及び受容体
Anti-Glutamate Decarboxylase Antibody, 65 kDa isoform, clone GAD-6 is an antibody against Glutamate Decarboxylase for use in IH & WB.
Immunohistochemistry: (≤ 1 μg/ml) Optimal working dilutions must be determined by end user.
Immunohystochemical Staining Procedures
The following procedure was developed to localize GAD in rat brain sections of cerebellum. Perform all steps at room temperature unless otherwise indicated. Where normal serum is indicated, use normal serum from the same species as the source of the secondary antibody.This procedure represents suggested guidelines for the use of anti-GAD. Fixation regimen, antibody concentrations, and incubation conditions for a given experimental system should be determined empirically.
1. Perfuse rats with 100 mM phosphate buffer, pH 7.4, containing 1% paraformaldehyde, 0.34% L-lysine, and 0.05% sodium m-periodate (1% PLP).
2. Postfix brains in 1% PLP for 1-2 hours. Longer fixation times may reduce labeling intensity.
3. Transfer brains to 100 mM phosphate buffer containing 30% sucrose, and gently agitate on a shaker platform at +4°C for 48-60 hours.
4. Using a sliding microtome, cut 30 mm sections of frozen cerebellum. As the sections are cut, collect them in a vial of cold 100 mM phosphate buffer.
5. Incubate sections in phosphate-buffered saline (PBS) containing 1.5% normal serum and 0.2% TritonX-100 for 30 minutes.
6. On a shaker platform, incubate sections with anti-GAD (diluted in PBS containing 1.5% normal serum and 0.2% Triton X-100 to a final antibody concentration of 1 mg/ml) for 12-36 hours at +4°C.
7. On a shaker platform, rinse sections eight times, 10-15 minutes per rinse, in PBS.
8. Detect with a standard secondary antibody detection system (Hsu et al., 1981; Falini & Taylor, 1983; Harlow & Lane, 1988; Taylor, 1978).
9. Mount sections, dehydrate, and apply coverslips.
Immunohystochemical Staining Procedures
The following procedure was developed to localize GAD in rat brain sections of cerebellum. Perform all steps at room temperature unless otherwise indicated. Where normal serum is indicated, use normal serum from the same species as the source of the secondary antibody.This procedure represents suggested guidelines for the use of anti-GAD. Fixation regimen, antibody concentrations, and incubation conditions for a given experimental system should be determined empirically.
1. Perfuse rats with 100 mM phosphate buffer, pH 7.4, containing 1% paraformaldehyde, 0.34% L-lysine, and 0.05% sodium m-periodate (1% PLP).
2. Postfix brains in 1% PLP for 1-2 hours. Longer fixation times may reduce labeling intensity.
3. Transfer brains to 100 mM phosphate buffer containing 30% sucrose, and gently agitate on a shaker platform at +4°C for 48-60 hours.
4. Using a sliding microtome, cut 30 mm sections of frozen cerebellum. As the sections are cut, collect them in a vial of cold 100 mM phosphate buffer.
5. Incubate sections in phosphate-buffered saline (PBS) containing 1.5% normal serum and 0.2% TritonX-100 for 30 minutes.
6. On a shaker platform, incubate sections with anti-GAD (diluted in PBS containing 1.5% normal serum and 0.2% Triton X-100 to a final antibody concentration of 1 mg/ml) for 12-36 hours at +4°C.
7. On a shaker platform, rinse sections eight times, 10-15 minutes per rinse, in PBS.
8. Detect with a standard secondary antibody detection system (Hsu et al., 1981; Falini & Taylor, 1983; Harlow & Lane, 1988; Taylor, 1978).
9. Mount sections, dehydrate, and apply coverslips.
ターゲットの説明
65 kDa
物理的形状
Ammonium sulfate precipitation and DEAE-cellulose chromatography
Format: Purified
Lyophilized. Dissolve contents of vial in 100 µL of sterile, distilled water. This results in a final antibody concentration of 1 mg/ml in 10 mM potassium phosphate, 70 mM sodium chloride, pH 7.4 containing no preservatives.
保管および安定性
Maintain for 1 year at -20°C from date of shipment. Aliquot to avoid repeated freezing and thawing. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
アナリシスノート
Control
Brain tissue
Brain tissue
その他情報
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
法的情報
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
免責事項
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
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シグナルワード
Warning
危険有害性情報
危険有害性の分類
Acute Tox. 4 Dermal - Acute Tox. 4 Inhalation - Acute Tox. 4 Oral - Aquatic Chronic 3
保管分類コード
13 - Non Combustible Solids
WGK
WGK 3
引火点(°F)
Not applicable
引火点(℃)
Not applicable
適用法令
試験研究用途を考慮した関連法令を主に挙げております。化学物質以外については、一部の情報のみ提供しています。 製品を安全かつ合法的に使用することは、使用者の義務です。最新情報により修正される場合があります。WEBの反映には時間を要することがあるため、適宜SDSをご参照ください。
毒物及び劇物取締法
毒物
労働安全衛生法名称等を表示すべき危険物及び有害物
名称等を表示すべき危険物及び有害物
労働安全衛生法名称等を通知すべき危険物及び有害物
名称等を通知すべき危険物及び有害物
Jan Code
MAB351R:
試験成績書（COA）
製品のロット番号・バッチ番号を入力して、試験成績書（COA） を検索できます。ロット番号・バッチ番号は、製品ラベルに「Lot」または「Batch」に続いて記載されています。
Diminished neurosteroid sensitivity of synaptic inhibition and altered location of the alpha4 subunit of GABA(A) receptors in an animal model of epilepsy.
Sun, C; Mtchedlishvili, Z; Erisir, A; Kapur, J
The Journal of Neuroscience null
Time-resolved fluorometric assay for detection of autoantibodies to glutamic acid decarboxylase (GAD65).
Matti Ankelo, Annette Westerlund-Karlsson, Jorma Ilonen, Mikael Knip, Kaisa Savola et al.
Clinical Chemistry null
Galanin receptor 1 is expressed in a subpopulation of glutamatergic interneurons in the dorsal horn of the rat spinal cord.
Marc Landry, Rabia Bouali-Benazzouz, Caroline Andre, Tie Jun Sten Shi, Claire Leger et al.
The Journal of Comparative Neurology null
Anatomy of glutamic acid decarboxylase immunoreactive neurons and axons in the rat medial geniculate body.
Winer, J A and Larue, D T
The Journal of Comparative Neurology, 278, 47-68 (1988)
Distribution of alpha1, alpha4, gamma2, and delta subunits of GABAA receptors in hippocampal granule cells.
Sun, C; Sieghart, W; Kapur, J
Brain Research null
資料
Human iPSC neural differentiation media and protocols used to generate neural stem cells, neurons and glial cell types.
ライフサイエンス、有機合成、材料科学、クロマトグラフィー、分析など、あらゆる分野の研究に経験のあるメンバーがおります。.
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