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Catalog Number | orb654392 |
---|---|
Category | Antibodies |
Description | Anti-GAD65/GAD2 Antibody (monoclonal, 7G2). Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. |
Species/Host | Mouse |
Clonality | Monoclonal |
Clone Number | 7G2 |
Tested applications | FC, ICC, IF, IHC, WB |
Reactivity | Human, Mouse, Rat |
Isotype | Mouse IgG2a |
Immunogen | A 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. |
Concentration | Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml. |
Form/Appearance | Lyophilized |
Conjugation | Unconjugated |
MW | 62 kDa |
UniProt ID | Q05329 |
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. |
Alternative names | chromosomal protein, Nonhistone, HMG4 antibody; Hi Read more... |
Note | For research use only |
Application notes | Western 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 Date | 12 months from date of receipt. |
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.
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.
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.
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.
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.
FC, ICC, IF, IHC, WB | |
Human, Mouse, Rat | |
Mouse | |
Monoclonal | |
iFluor647 |
FC, ICC, IF, IHC, WB | |
Human, Mouse, Rat | |
Mouse | |
Monoclonal | |
PE |
FC, ICC, IF, IHC, WB | |
Human, Mouse, Rat | |
Mouse | |
Monoclonal | |
APC |
FC, ICC, IF, IHC, WB | |
Human, Mouse, Rat | |
Mouse | |
Monoclonal | |
HRP |
FC, ICC, IF, IHC, WB | |
Human, Mouse, Rat | |
Mouse | |
Monoclonal | |
FITC |