Anti-GAD65 Antibody, clone 3B23 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1091 - recombinant, expressed in HEK 293 cells
Synonym: 65 kDa glutamic acid decarboxylase; EC:4.1.1.15; GAD-65; Glutamate decarboxylase 2; Glutamate decarboxylase 65 kDa isoform
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human cerebral cortex (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:10 dilution of Cat. No. ZRB1091, Anti-GAD65, clone 3B23 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in subsets of neurons and neuronal processes of human cerebral cortex tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human cerebral cortex (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:10 dilution of Cat. No. ZRB1091, Anti-GAD65, clone 3B23 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in subsets of neurons and neuronal processes of human cerebral cortex tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human cerebral cortex (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:10 dilution of Cat. No. ZRB1091, Anti-GAD65, clone 3B23 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in subsets of neurons and neuronal processes of human cerebral cortex tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) rat cerebellum (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1091, Anti-GAD65, clone 3B23 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in granular and molecular layers of rat cerebellum tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) rat cerebellum (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1091, Anti-GAD65, clone 3B23 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in granular and molecular layers of rat cerebellum tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) rat cerebellum (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1091, Anti-GAD65, clone 3B23 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in granular and molecular layers of rat cerebellum tissue sections.
Immunofluorescence Enhanced Validation-Recombinant Antibody Technology Confocal fluorescent analysis of rat cerebellum tissue sections was performed using a 1:100 dilution of Cat. No. ZRB1091, Anti-GAD65, clone 3B23 ZooMAb® Rabbit Monoclonal and visualized with a secondary antibody conjugated to Alexa Fluor® 488 (Green). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasmic membrane.
Immunofluorescence Enhanced Validation-Recombinant Antibody Technology Confocal fluorescent analysis of rat cerebellum tissue sections was performed using a 1:100 dilution of Cat. No. ZRB1091, Anti-GAD65, clone 3B23 ZooMAb® Rabbit Monoclonal and visualized with a secondary antibody conjugated to Alexa Fluor® 488 (Green). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasmic membrane.
Immunofluorescence Enhanced Validation-Recombinant Antibody Technology Confocal fluorescent analysis of rat cerebellum tissue sections was performed using a 1:100 dilution of Cat. No. ZRB1091, Anti-GAD65, clone 3B23 ZooMAb® Rabbit Monoclonal and visualized with a secondary antibody conjugated to Alexa Fluor® 488 (Green). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasmic membrane.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of Neuro-2a cell line were performed using a 1:100 ilution of Cat. No. ZRB1091, Anti-GAD65, clone 3B23 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488. Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of Neuro-2a cell line were performed using a 1:100 ilution of Cat. No. ZRB1091, Anti-GAD65, clone 3B23 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488. Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of Neuro-2a cell line were performed using a 1:100 ilution of Cat. No. ZRB1091, Anti-GAD65, clone 3B23 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488. Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
Western Blotting Enhanced Validation-Recombinant Antibody Technology Human Brain (Lane 1), Mouse Brain (Lane 2), and Rat Brain (Lane 3) tissue lysates were probed with Cat. No. ZRB1091, Anti-GAD65, clone 3B23 ZooMAb® Rabbit Monoclonal (1:1,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates GAD65 (~58 kDa).
Affinity Binding Assay Enhanced Validation-Recombinant Antibody Technology 100 μg/mL of non-specific control peptide (Run 1) or GAD65 peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1091, Anti-GAD65, clone 3B23 ZooMAb® Rabbit Monoclonal. The binding of this antibody to GAD65 peptide displayed a KD value of 1 x 10-12 whereas its binding to non-specific peptide displayed a KD of 6.8 x 10-5.
12 Principles of Green Chemistry: Principle 1—Waste Prevention. This product has a smaller environmental footprint through reduction of waste in the manufacturing process.
12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.
Chemical products should be designed to affect their desired function while minimizing their toxicity.
12 Principles of Green Chemistry: Principle 6—Design for Energy Efficiency. This product was re-engineered to utilize less energy in the manufacturing process.
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| biological source | rabbit |
| clone | 3B23, monoclonal |
| recombinant monoclonal | |
| conjugate | unconjugated |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| form | lyophilized |
| greener alternative category | Aligned |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry . |
| mol wt | calculated mol wt 65.41 kDa |
| observed mol wt ~58 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | rat, mouse, human |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| immunocytochemistry: suitable | |
| immunofluorescence: suitable | |
| immunohistochemistry: suitable | |
| western blot: suitable | |
| UniProt accession no. | Q05329  |
| Application: | Anti-GAD65, clone 3B23 ZooMAb, Cat. No. ZRB1091, is a recombinant Rabbit monoclonal antibody that targets GAD65 and is tested for use in Affinity Binding Assay, Immunocytochemistry, Immunofluorescence, Immunohistochemistry, and Western Blotting. |
| Application: | Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected GAD65 human and mouse brain tissue lysates. Immunofluorescence Analysis: A 1:100 dilution from a representative lot detected GAD65 in rat cerebellum tissue sections. Affinity Binding Assay:: A representative lot of this antibody bound human GAD65 peptide with a KD of 1 x 10-12 in an affinity binding assay. Immunohistochemistry (Paraffin) Analysis: A 1:10-100 dilution from a representative lot detected GAD65 in human brain and rat brain tissue sections. Immunocytochemistry Analysis: A 1:100 dilution from a representativelot detected GAD65 in Neuro-2a cells. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| Disclaimer: | 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. |
| General description: | We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information. |
| General description: | ZooMAb antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb antibodies are reliably available and ready to ship when you need them. Learn more about ZooMAb here. |
| Immunogen: | KLH-conjugated linear peptide corresponding to 17 amino acids from the C-terminal region of human Glutamate decarboxylase 2 (GAD65). |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL. |
| Reconstitution: | 30 μg/mL after reconstitution at 25 μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 2B23 is a ZooMAb rabbit recombinant monoclonal antibody that specifically detects Glutamate decarboxylase 2 (GAD65). It targets an epitope within 17 amino acids from the C-terminal region. |
| Storage and Stability: | Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws. |
| Target description: | Glutamate decarboxylase 2 (UniProt: Q05329; also known as EC:4.1.1.15, 65 kDa glutamic acid decarboxylase, GAD-65, Glutamate decarboxylase 65 kDa isoform) is encoded by the GAD2 (GAD65) gene (Gene ID: 2572) in human. GAD-65 a homodimeric peripheral membrane protein that is associated to cytoplasmic vesicles. It is produced primarily by pancreatic islet cells. It is expressed in human β cells; however, some expression has also been observed in and cells of islets. It is shown to be a major target autoantigen in type 1A diabetes. In neurons, it is present in cytosolic leaflet of Golgi membranes and presynaptic clusters. It is responsible for the conversion of glutamic acid to gamma-aminobutyric acid (GABA), the major inhibitory transmitter in higher brain regions. Two molecular forms of GAD have been described (65 kDa and 67 kDa) that display 64% amino acid homology and both forms are expressed in the CNS, pancreatic islet cells, testis, oviduct, and ovary. Between the two isoforms, GAD67 is responsible for over 90% of basal GABA synthesis and is produced at limiting levels in the brain. This ZooMAb recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. |
| WGK Germany | WGK 1 |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |