Product Info Summary
SKU: | A01758-2 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Rabbit |
Application: | ELISA, Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-METTL3 Antibody Picoband®
View all Methyltransferase like 3 Antibodies
SKU/Catalog Number
A01758-2
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-METTL3 Antibody Picoband® catalog # A01758-2. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
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 # A01758-2)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.005mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human METTL3 recombinant protein (Position: E71-L580).
*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
A01758-2 is reactive to METTL3 in Human, Mouse, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500μg/ml.
Observed Molecular Weight
64 kDa
Calculated molecular weight
64.616kDa
Background of Methyltransferase like 3
N6-adenosine-methyltransferase 70 kDa subunit (METTL3) is an enzyme that in humans is encoded by the METTL3 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.
Application & Images
Applications
A01758-2 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
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, 0.25-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, Human
Positive Control
WB: human PC-3 whole cell, human HEPG2 whole cell, human A549 whole cell, human HEK293 whole cell, human HELA whole cell, human CACO-2 whole cell, rat testis tissue, mouse testis tissue
IHC: human esophageal squamous carcinoma tissue, human tonsil tissue, human placenta tissue, rat epididymis tissue
ICC/IF: PC-3 cell
FCM: HL-60 cell
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of METTL3 using anti-METTL3 antibody (A01758-2).
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 PC-3 whole cell lysates,
Lane 2: human HEPG2 whole cell lysates,
Lane 3: human A549 whole cell lysates,
Lane 4: human HEK293 whole cell lysates,
Lane 5: human HELA whole cell lysates,
Lane 6: human CACO-2 whole cell lysates,
Lane 7: rat testis tissue lysates,
Lane 8: mouse testis 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 rabbit anti-METTL3 antigen affinity purified polyclonal antibody (Catalog # A01758-21) 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 64KD. The expected band size for METTL3 is at 64KD.
Click image to see more details
Figure 2. IHC analysis of METTL3 using anti-METTL3 antibody (A01758-2).
METTL3 was detected in paraffin-embedded section of human esophageal squamous carcinoma 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 2μg/ml rabbit anti-METTL3 Antibody (A01758-2) 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) (Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 3 IHC analysis of METTL3 using anti-METTL3 antibody (A01758-2).
METTL3 was detected in paraffin-embedded section of human tonsil 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 2μg/ml rabbit anti-METTL3 Antibody (A01758-2) 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) (Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 4 IHC analysis of METTL3 using anti-METTL3 antibody (A01758-2).
METTL3 was detected in paraffin-embedded section of human placenta 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 2μg/ml rabbit anti-METTL3 Antibody (A01758-2) 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) (Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 5. IF analysis of METTL3 using anti-METTL3 antibody (A01758-2).
METTL3 was detected in immunocytochemical section of PC-3 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-METTL3 Antibody (A01758-2) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) 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.
Click image to see more details
Figure 6. Flow Cytometry analysis of HL-60 cells using anti-METTL3 antibody (A01758-2).
Overlay histogram showing HL-60 cells stained with A01758-2 (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 (A01758-2, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Click image to see more details
Figure 7. IHC analysis of METTL3 using anti-METTL3 antibody (A01758-2).
METTL3 was detected in a paraffin-embedded section of rat epididymis tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-METTL3 Antibody (A01758-2) 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) (Catalog # SA1022) with DAB as the chromogen.
Protein Target Info & Infographic
Gene/Protein Information For METTL3 (Source: Uniprot.org, NCBI)
Gene Name
METTL3
Full Name
N6-adenosine-methyltransferase subunit METTL3
Weight
64.616kDa
Superfamily
MT-A70-like family
Alternative Names
N6-adenosine-methyltransferase catalytic subunit 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
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on METTL3 , check out the METTL3 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
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].
Specific Publications For Anti-METTL3 Antibody Picoband® (A01758-2)
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