Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [1]
- Immunocytochemistry [3]
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- Product number
- LS-C312273 - Provider product page
- Provider
- LSBio
- Product name
- GAD1 / GAD67 Antibody (aa10-24) LS-C312273
- Antibody type
- Polyclonal
- Description
- Immunogen affinity purified
- Reactivity
- Human, Mouse, Rat, Bovine, Guinea Pig, Rabbit, Simian
- Host
- Rabbit
- Storage
- At -20°C for 1 year. After reconstitution, at 4°C for 1 month. It can also be aliquotted and stored frozen at -20°C for a longer time. Avoid freeze-thaw cycles.
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Supportive validation
- Submitted by
- LSBio (provider)
- Enhanced method
- Genetic validation
- Main image
- Experimental details
- Western blot analysis of GAD67 using anti-GAD67 antibody. 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 Lysate, Lane 2: Rat Testis Tissue Lysate, Lane 3: MCF-7 Whole Cell Lysate, Lane 4: MM231 Whole Cell Lysate, Lane 5: HELA Whole Cell Lysate, Lane 6: SMMC Whole Cell Lysate, Lane 7: COLO320 Whole Cell Lysate. 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 rabbit anti-GAD67 antigen affinity purified polyclonal antibody 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-rabbit 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 with Tanon 5200 system. A specific band was detected for GAD67 at approximately 67KD. The expected band size for GAD67 is at 67KD.
Supportive validation
- Submitted by
- LSBio (provider)
- Enhanced method
- Genetic validation
- Main image
- Experimental details
- ICC analysis of GAD67 using anti-GAD67 antibody. GAD67 was detected in immunocytochemical section of A431 cells. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 3µg/ml rabbit anti-GAD67 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
- Submitted by
- LSBio (provider)
- Main image
- Experimental details
- ICC analysis of GAD67 using anti-GAD67 antibody. GAD67 was detected in immunocytochemical section of A431 cells. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 3µg/ml rabbit anti-GAD67 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
- Submitted by
- LSBio (provider)
- Main image
- Experimental details
- ICC analysis of GAD67 using anti-GAD67 antibody. GAD67 was detected in immunocytochemical section of A431 cells. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 3µg/ml rabbit anti-GAD67 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.