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GAD65/GAD2 Antibody (monoclonal, 7G2)

Catalog Number: orb654392

DispatchUsually dispatched within 5-10 working days
$ 210.00
Catalog Numberorb654392
CategoryAntibodies
DescriptionAnti-GAD65/GAD2 Antibody (monoclonal, 7G2). Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
Species/HostMouse
ClonalityMonoclonal
Clone Number7G2
Tested applicationsFC, ICC, IF, IHC, WB
ReactivityHuman, Mouse, Rat
IsotypeMouse IgG2a
ImmunogenA synthetic peptide corresponding to a sequence at the N-terminus of human GAD65, different from the related mouse and rat sequences by one amino acid.
ConcentrationAdding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
Form/AppearanceLyophilized
ConjugationUnconjugated
MW62 kDa
UniProt IDQ05329
StorageMaintain 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.
Alternative nameschromosomal protein, Nonhistone, HMG4 antibody; Hi
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NoteFor research use only
Application notesWestern blot, 0.1-0.5μg/ml, Mouse, RatImmunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, HumanImmunocytochemistry/Immunofluorescence, 4μg/ml, Human Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
Expiration Date12 months from date of receipt.
GAD65/GAD2 Antibody (monoclonal, 7G2)

Western blot analysis of GAD65/GAD2 using anti-GAD65/GAD2 antibody (orb654392). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: rat brain tissue lysates, Lane 2: mouse brain tissue lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-GAD65/GAD2 antigen affinity purified monoclonal antibody (Catalog # orb654392) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # orb90502) with Tanon 5200 system. A specific band was detected for GAD65/GAD2 at approximately 62KD. The expected band size for GAD65/GAD2 is at 62KD.

GAD65/GAD2 Antibody (monoclonal, 7G2)

IHC analysis of GAD65/GAD2 using anti-GAD65/GAD2 antibody (orb654392). GAD65/GAD2 was detected in paraffin-embedded section of human glioma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-GAD65/GAD2 Antibody (orb654392) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90443) with DAB as the chromogen.

GAD65/GAD2 Antibody (monoclonal, 7G2)

IF analysis of GAD65/GAD2 using anti-GAD65/GAD2 antibody (orb654392). GAD65/GAD2 was detected in immunocytochemical section of HeLa cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (orb90553) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 4μg/mL mouse anti-GAD65/GAD2 Antibody (orb654392) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

GAD65/GAD2 Antibody (monoclonal, 7G2)

Flow Cytometry analysis of U20S cells using anti-GAD65/GAD2 antibody (orb654392). Overlay histogram showing U20S cells stained with orb654392 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-GAD65/GAD2 Antibody (orb654392, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

GAD65/GAD2 Antibody (monoclonal, 7G2)

Flow Cytometry analysis of 293T cells using anti-GAD65/GAD2 antibody (orb654392). Overlay histogram showing 293T cells stained with orb654392 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-GAD65/GAD2 Antibody (orb654392, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.