GAD65 Rabbit Polyclonal Antibody

SKU: orb10682

Description

GAD65 Rabbit Polyclonal Antibody

Research Area

Amino Acid Metabolism, Glutamate, Metabolism, Neuroscience

Images & Validation

• Tested Applications: FC, ICC, WB
• Dilution range: WB=1:500-2000, ICC/IF=1:100-500, Flow-Cyt=1ug/Test
• Reactivity: Human, Mouse, Rat
• Predicted Reactivity: Bovine, Canine, Gallus, Porcine

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human GAD65 (501-585/585aa)
• Target: GAD2
• Molecular Weight: 65 kDa
• Purification: Affinity purified by Protein A
• Conjugation: Unconjugated

Storage & Handling

• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Form/Appearance: Liquid
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Concentration: 1mg/ml
• Disclaimer: For research use only

Alternative Names

65 kDa glutamic acid decarboxylase; DCE 2; DCE2; GAD 2; GAD 65; GAD-2; GAD-65; GAD 65, GAD2; Glutamate Decarboxylase 2(pancreatic islets and brain 65kDa); Glutamate Decarboxylase 2; Glutamate Decarboxylase 65; Glutamate decarboxylase 65 kDa isoform; Glutamic Acid Decarboxylase 2; Glutamic Acid Decarboxylase 65.

Blank control (Black line): Molt4 (Black). Primary Antibody (green line): Rabbit Anti-GAD65 antibody (orb10682), Dilution: 3 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF647, Dilution: 3 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with PBST for 20 min at room temperature. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Molt-4 cells were fixed with 4% PFA for 10 min at room temperature, permeabilized with 0.1% PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with GAD65 Antibody (orb10682) at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).

Sample: Lane 1: Mouse Cerebrum tissue lysates, Lane 2: Mouse Cerebellum tissue lysates, Lane 3: Mouse Hippocampus tissue lysates, Lane 4: Rat Cerebrum tissue lysates, Lane 5: Rat Cerebellum tissue lysates, Lane 6: Rat Hippocampus tissue lysates, Lane 7: Human SH-SY5Y cell lysates, Lane 8: Human U87MG cell lysates, Lane 9: Human HeLa cell lysates, Lane 10: Human Jurkat cell lysates, Primary: Anti-GAD65 (orb10682) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 65 kDa, Observed band size: 62 kDa.

SHSY5Y cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (GAD65) polyclonal Antibody, Unconjugated (orb10682) 1:100, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

Quick Database Links

Gene Symbol

GAD2

UniProt

Q05329

References

Janzadeh, Atousa et al. The effect of chondroitinase ABC and photobiomodulation therapy on neuropathic pain after spinal cord injury in adult male rats Physiol Behav, 227, 113141 (2020)

PubMed: 32818524