GAD Antikörper (C-Term)

Produktnummer ABIN3184725

Dieser Anti-GAD Antikörper ist ein Kaninchen Polyklonal Antikörper zur Detektion von GAD in WB, IHC und ELISA. Geeignet für Human, Maus und Ratte. Unabhängig validiert für die Verwendung in Immunohistochemistry.

Kurzübersicht für GAD Antikörper (C-Term) (ABIN3184725)

Target: GAD (GAD1) (Glutamate Decarboxylase 1 (Brain, 67kDa) (GAD1))

Reaktivität: Human, Maus, Ratte

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser GAD Antikörper ist unkonjugiert

Applikation: Western Blotting (WB), Immunohistochemistry (IHC), ELISA

Produktdetails anti-GAD Antikörper

Bindungsspezifität: C-Term

Spezifität: GAD67 Polyclonal Antibody detects endogenous levels of GAD67 protein.

Produktmerkmale: Rabbit Polyclonal to GAD67.

Aufreinigung: The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen.

Immunogen: Synthesized peptide derived from the C-terminal region of human GAD67.

Isotyp: IgG

Anwendungsinformationen

Applikationshinweise: WB 1:500-1:2000, IHC 1:100-1:300, ELISA 1:10000,

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (IHC)

Validierung #103754 (Immunohistochemistry)

by: Prof. Merighi, Laboratory of Neurobiology, Department of Veterinary Sciences, University of Turin

No.: #103754

Datum: 12.12.2019

Antigen: GAD1

Validierte Anwendung: Immunohistochemistry

Positivkontrolle:

6 µm glass-mounted microtome sections from adult (24 months) mouse brain

Negativkontrolle:

Slices incubated overnight with the blocking solution only and then processed with the secondary antibody

Bewertung:

Passed. The anti-GAD antibody ABIN3184725 specifically recognized GAD1 at a dilution of 1:200.

Validierungsbilder

A and B. Detection of GAD1+ neurons in the cortex using ABIN3184725.

Protokoll

Primärantikörper: ABIN3184725

Sekundärantikörper: goat anti-rabbit AF594-conjugated antibody (Thermo Fisher Scientific, A11029, lot 1942237)

Full Protocol:

• Perfuse mouse with 4% paraformaldehyde in 0.1 M phosphate buffer (PB) pH7.4 and post-fix spinal cord and brain samples in the same fixative for 2 h at RT.
• Wash samples in PBS, dehydrate, and embed samples in paraffin wax.
• Cut spinal cord and brain samples with a microtome in 6 µm-thick sections and mount sections on glass slides.
• After paraffin removal, block sections in PBS containing 1% albumin from chicken egg white (Sigma, A5378) and 0.3% Triton-X-100 (BioRad, 161-0407, lot 00583) 1 h at RT.
• Incubate sections with primary rabbit anti-GAD antibody (antibodies-online, ABIN3184725, lot 31965) diluted 1:200 in blocking solution ON at RT.
• Wash sections 3x 5 min in 0.01 M PBS.
• Incubate sections with secondary goat anti-rabbit AF594-conjugated antibody (Thermo Fisher Scientific, A11029, lot 1942237) diluted 1:200 in PBS for 1 h at RT.
• Wash sections 3x 5 min in 0.01 M PBS.
• Mount specimens in Fluoroshield (Sigma, F6182, lot MKCB0153V).
• Acquire images with Leica DM 6000B fluorescence microscope equipped with a digital camera at 40x magnification. Maintain Parameters for image acquisition (Exposure 2.2 s, Gain 2.3x, Saturation 1.5, Gamma 1.10) unchanged for all images.

Anmerkungen:

Dilutions of 1:50, 1:100, and 1:500 were also tested but 1:200 gave the best results.

Handhabung

Format: Liquid

Konzentration: 1 mg/mL

Buffer: Liquid in PBS containing 50 % glycerol, 0.5 % BSA and 0.02 % sodium azide.

Konservierungsmittel: Sodium azide

Vorsichtsmaßnahmen: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Handhabung: Avoid repeated freeze/thaw cycles.

Lagerung: -20 °C

Informationen zur Lagerung: Store at -20°C.

Details zu GAD

Target: GAD (GAD1) (Glutamate Decarboxylase 1 (Brain, 67kDa) (GAD1))

Andere Bezeichnung: GAD67

Molekulargewicht: 67 kDa

Gen-ID: 2571

UniProt: Q99259