GAD antibody (C-Term)

Catalog No. ABIN3184725

The Rabbit Polyclonal anti-GAD antibody (ABIN3184725) specifically detects GAD in WB, IHC and ELISA. The antibody is reactive with Human, Mouse and Rat samples. Independently validated for use in Immunohistochemistry.

Quick Overview for GAD antibody (C-Term) (ABIN3184725)

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

Reactivity: Human, Mouse, Rat

Host: Rabbit

Clonality: Polyclonal

Conjugate: This GAD antibody is un-conjugated

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

Product Details anti-GAD Antibody

Binding Specificity: C-Term

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

Characteristics: Rabbit Polyclonal to GAD67.

Purification: 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.

Isotype: IgG

Application Details

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

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #103754 (Immunohistochemistry)

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

No.: #103754

Date: 12/12/2019

Antigen: GAD1

Method validated: Immunohistochemistry

Positive Control:

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

Negative Control:

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

Notes:

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

Validation Images

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

Full Methods

Primary Antibody: ABIN3184725

Secondary Antibody: 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.

Experimental Notes:

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

Handling

Format: Liquid

Concentration: 1 mg/mL

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

Preservative: Sodium azide

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

Handling Advice: Avoid repeated freeze/thaw cycles.

Storage: -20 °C

Storage Comment: Store at -20°C.

Target Details for GAD

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

Alternative Name: GAD67

Molecular Weight: 67 kDa

Gene ID: 2571

UniProt: Q99259