MAB351R
Anti-GAD2(GAD67) Antibody
CHEMICON®, mouse monoclonal, GAD-6
Synonim(y):
GAD65
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About This Item
Kod UNSPSC:
12352203
eCl@ss:
32160702
NACRES:
NA.41
Polecane produkty
product name
Anti-Glutamate Decarboxylase Antibody, 65 kDa isoform, clone GAD-6, clone GAD-6, Chemicon®, from mouse
pochodzenie biologiczne
mouse
Poziom jakości
forma przeciwciała
purified immunoglobulin
rodzaj przeciwciała
primary antibodies
klon
GAD-6, monoclonal
reaktywność gatunkowa
human, rat
producent / nazwa handlowa
Chemicon®
metody
immunohistochemistry: suitable
western blot: suitable
izotyp
IgG2a
numer dostępu NCBI
numer dostępu UniProt
Warunki transportu
dry ice
docelowa modyfikacja potranslacyjna
unmodified
informacje o genach
human ... GAD2(2572)
Opis ogólny
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.
Specyficzność
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).
Immunogen
Purified rat brain GAD.
Zastosowanie
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.
Research Category
Neuroscience
Neuroscience
Research Sub Category
Neurotransmitters & Receptors
Neurotransmitters & Receptors
Opis wartości docelowych
65 kDa
Postać fizyczna
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.
Przechowywanie i stabilność
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.
Komentarz do analizy
Control
Brain tissue
Brain tissue
Inne uwagi
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Informacje prawne
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Oświadczenie o zrzeczeniu się odpowiedzialności
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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Hasło ostrzegawcze
Warning
Zwroty wskazujące rodzaj zagrożenia
Zwroty wskazujące środki ostrożności
Klasyfikacja zagrożeń
Acute Tox. 4 Dermal - Acute Tox. 4 Inhalation - Acute Tox. 4 Oral - Aquatic Chronic 3
Kod klasy składowania
13 - Non Combustible Solids
Klasa zagrożenia wodnego (WGK)
WGK 3
Temperatura zapłonu (°F)
Not applicable
Temperatura zapłonu (°C)
Not applicable
Certyfikaty analizy (CoA)
Poszukaj Certyfikaty analizy (CoA), wpisując numer partii/serii produktów. Numery serii i partii można znaleźć na etykiecie produktu po słowach „seria” lub „partia”.
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Dokumenty związane z niedawno zakupionymi produktami zostały zamieszczone w Bibliotece dokumentów.
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
Produkty
Human iPSC neural differentiation media and protocols used to generate neural stem cells, neurons and glial cell types.
Podłoża do różnicowania neuronalnego ludzkich iPSC i protokoły stosowane do generowania neuronalnych komórek macierzystych, neuronów i komórek glejowych.
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