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Western blot
ELISA
ImmunocytochemistryAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
- Immunocytochemistry [2]
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- Product number
- LS-C756516 - Provider product page
- Provider
- LSBio
- Product name
- METTL3 Antibody LS-C756516
- Antibody type
- Polyclonal
- Description
- Immunogen affinity purified
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Storage
- After reconstitution, may be stored at 4°C for 1 month. For long-term storage and to avoid freeze-thaw cycles, aliquot and store at -20°C.
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Enhanced validation
- Submitted by
- LSBio (provider)
- Enhanced method
- Genetic validation
- Main image
- Experimental details
- Western blot analysis of METTL3 using anti-METTL3 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: human placenta tissue lysates,Lane 2: human HepG2 whole cell lysate,Lane 3: human K562 whole cell lysate,Lane 4: human Hela whole cell lysate,Lane 5: human Caco-2 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-METTL3 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 METTL3 at approximately 64KD. The expected band size for METTL3 is at 64KD.
Supportive validation
- Submitted by
- LSBio (provider)
- Enhanced method
- Genetic validation
- Main image
- Experimental details
- ICC analysis of METTL3 using anti-METTL3 antibody. METTL3 was detected in immunocytochemical section of A549 cell. 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 1µg/ml rabbit anti-METTL3 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 METTL3 using anti-METTL3 antibody. METTL3 was detected in immunocytochemical section of A549 cell. 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 1µg/ml rabbit anti-METTL3 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.
