GAD antibody (C-Term)

Catalog No. ABIN3184725

This anti-GAD antibody is a Rabbit Polyclonal antibody detecting GAD in WB, IHC and ELISA. Suitable for Human, Mouse and Rat. 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