추천 제품
제품명
Anti-Glutamate Decarboxylase Antibody, 65 kDa isoform, clone GAD-6, clone GAD-6, Chemicon®, from mouse
생물학적 소스
mouse
Quality Level
항체 형태
purified immunoglobulin
항체 생산 유형
primary antibodies
클론
GAD-6, monoclonal
종 반응성
rat, human
제조업체/상표
Chemicon®
기술
immunohistochemistry: suitable
western blot: suitable
동형
IgG2a
NCBI 수납 번호
UniProt 수납 번호
배송 상태
dry ice
타겟 번역 후 변형
unmodified
유전자 정보
human ... GAD2(2572)
일반 설명
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 (65 kDa and 67 kDa, 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 (585 a.a.), encoded on human chromosome 10, and is responsible for vesicular GABA production. GAD67 is cytoplasmic (594 a.a.), 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.
특이성
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).
면역원
Purified rat brain glutamic acid decarboxylase.
애플리케이션
Anti-GAD Antibody. 65 kDa isoform, clone GAD-6 detects level of Glutamate Decarboxylase & has been published & validated for use in IH & WB.
Immunohistochemistry: ( 1 μg/mL * See protocol)
Western blot
Optimal working dilutions must be determined by end user.
APPLICATION NOTES FOR MAB351
IMMUNOHISTOCHEMISTRY
1) Perfuse rats with 100 mM phosphate buffer, pH 7.4 containing 1% paraformaldehyde, 0.34% L-lysine and 0.05% m-periodate (1% PLP).
2) Postfix brains in 1% PLP for 1-2 hours.
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 PBS containing 1.5% normal serum and 0.2% Triton X-100 for 30 minutes.
6) On a shaker platform, incubate sections with MAB351 (diluted 1 μg/mL in PBS containing 1.5% normal serum and 0.2% Triton X-100) 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 standard secondary antibody detection system (PAP, ABC, etc.).
9) Mount sections, dehydrate, and apply coverslips.
Western blot
Optimal working dilutions must be determined by end user.
APPLICATION NOTES FOR MAB351
IMMUNOHISTOCHEMISTRY
1) Perfuse rats with 100 mM phosphate buffer, pH 7.4 containing 1% paraformaldehyde, 0.34% L-lysine and 0.05% m-periodate (1% PLP).
2) Postfix brains in 1% PLP for 1-2 hours.
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 PBS containing 1.5% normal serum and 0.2% Triton X-100 for 30 minutes.
6) On a shaker platform, incubate sections with MAB351 (diluted 1 μg/mL in PBS containing 1.5% normal serum and 0.2% Triton X-100) 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 standard secondary antibody detection system (PAP, ABC, etc.).
9) Mount sections, dehydrate, and apply coverslips.
Research Category
Neuroscience
Neuroscience
Research Sub Category
Neurotransmitters & Receptors
Neurotransmitters & Receptors
표적 설명
65 kDa
물리적 형태
Format: Purified
Protein A purified
Purified immunoglobulin. Lyophilized from 10 mM potassium phosphate, 70 mM sodium chloride, pH 7.4, 0.02% sodium azide. Reconstitute with 100 μL of sterile water (1 mg/mL).
저장 및 안정성
Maintain lyophilized material at -20°C for up to 12 months. After reconstitution maintain frozen at -20°C in undiluted aliquots for up to 6 months. Avoid repeated freeze/thaw cycles.
분석 메모
Control
Rat brain tissue
human brain lysate
Rat brain tissue
human brain lysate
기타 정보
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
법적 정보
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
면책조항
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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신호어
Warning
유해 및 위험 성명서
Hazard Classifications
Acute Tox. 4 Dermal - Acute Tox. 4 Inhalation - Acute Tox. 4 Oral - Aquatic Chronic 3
Storage Class Code
13 - Non Combustible Solids
WGK
WGK 3
Flash Point (°F)
Not applicable
Flash Point (°C)
Not applicable
시험 성적서(COA)
제품의 로트/배치 번호를 입력하여 시험 성적서(COA)을 검색하십시오. 로트 및 배치 번호는 제품 라벨에 있는 ‘로트’ 또는 ‘배치’라는 용어 뒤에서 찾을 수 있습니다.
Differential activation of neuronal cell types in the basolateral amygdala by corticotropin releasing factor.
Rostkowski, AB; Leitermann, RJ; Urban, JH
Neuropeptides null
Synaptic reorganisation of the medial amygdala during puberty.
Cooke BM
Journal of Neuroendocrinology null
Thalamocortical dysfunction and thalamic injury after asphyxial cardiac arrest in developing rats.
Shoykhet, M; Simons, DJ; Alexander, H; Hosler, C; Kochanek, PM; Clark, RS
The Journal of Neuroscience null
Quantitative effects produced by modifications of neuronal activity on the size of GABAA receptor clusters in hippocampal slice cultures.
Serge Marty, Rosine Wehrle, Jean-Marc Fritschy, Constantino Sotelo
The European Journal of Neuroscience null
Quantitative analysis of ER alpha and GAD colocalization in the hippocampus of the adult female rat.
S A Hart, J D Patton, C S Woolley, S A Hart, J D Patton, C S Woolley
The Journal of Comparative Neurology null
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