Anti-METTL3 Antibody Picoband™

Product Name

Anti-METTL3 Antibody Picoband™

View all Methyltransferase like 3 Antibodies

SKU/Catalog Number

A01758-1

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-METTL3/METTL3 Antibody Picoband™ catalog # A01758-1. Tested in ELISA, Flow Cytometry, WB applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-METTL3 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A01758-1)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E. coli-derived human METTL3 recombinant protein (Position: Q423-D571).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A01758-1 is reactive to Mettl3 in Human, Mouse, Rat

Applications

A01758-1 is guaranteed for ELISA, Flow Cytometry, WB Boster Guarantee

Observed Molecular Weight

64 kDa

Calculated molecular weight

Background of Methyltransferase like 3

N6-adenosine-methyltransferase 70 kDa subunit (METTL3) is an enzyme that in humans is encoded by theMETTL3 gene. It is mapped to 14q11.2. This gene encodes the 70 kDa subunit of MT-A which is part of N6-adenosine-methyltransferase. This enzyme is involved in the posttranscriptional methylation of internal adenosine residues in eukaryotic mRNAs, forming N6-methyladenosine.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconstitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.1-0.5μg/ml
Flow Cytometry (Fixed), 1-3 μg/1x10⁶ cells
Direct ELISA, 0.1-0.5μg/ml

Validation Images & Assay Conditions

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Figure 2. Flow Cytometry analysis of HEPA1-6 cells using anti-METTL3 antibody (A01758-1).
Overlay histogram showing HEPA1-6 cells stained with A01758-1 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-METTL3 Antibody (A01758-1, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

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Figure 1. Western blot analysis of METTL3 using anti-METTL3 antibody (A01758-1).
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 30 ug of sample under reducing conditions.
Lane 1: human HepG2 whole cell lysates,
Lane 2: human PC-3 whole cell lysates,
Lane 3: human A549 whole cell lysates,
Lane 4: human Hacat whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: rat testis tissue lysates,
Lane 7: mouse brain tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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 (Catalog # A01758-1) 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 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for METTL3 at approximately 64 kDa. The expected band size for METTL3 is at 64 kDa.

Gene/Protein Information For Mettl3 (Source: Uniprot.org, NCBI)

Alternative Names

adoMet-binding subunit of the human mRNA (N6-adenosine)-methyltransferase; EC 2.1.1.62; IME4; M6A; methyltransferase like 3; Methyltransferase-like protein 3; MGC4336; mRNA m(6)A methyltransferase; MTA70; MT-A70; N6-adenosine-methyltransferase 70 kDa subunit; Spo8 Mettl3|2310024F18Rik, M6, M6A, S, Spo8|methyltransferase like 3|N6-adenosine-methyltransferase subunit METTL3|MT-A70|N6-adenosine-methyltransferase 70 kDa subunit|m6a methyltransferase|methyltransferase-like protein 3

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In this infographic, you will see the following information for Mettl3: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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