Recombinant Anti-METTL3 antibody [EPR18810] (ab195352)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18810] to METTL3
- Suitable for: Flow Cyt, WB, IHC-P, ICC/IF, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-METTL3 antibody [EPR18810]
See all METTL3 primary antibodies -
Description
Rabbit monoclonal [EPR18810] to METTL3 -
Host species
Rabbit -
Tested Applications & Species
Application Species Flow Cyt HumanICC/IF HumanIHC-P MouseRatHumanIP HumanWB MouseRatHuman -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Raji, HeLa, HEK-293, Jurkat, NCCIT, F9, Neuro-2a, LLC1, C6, RAW 264.7, PC-12 and NIH\3T3 cell lysates; human thymus lysate, mouse brain, spleen and heart lysates; rat brain lysate. IHC-P: Human bladder cancer, mouse testis and rat testis tissues. ICC/IF: HCT116 and HeLa cells. IP: HeLa cell lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18810 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab195352 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
Application | Species |
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Flow Cyt |
Human
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ICC/IF |
Human
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IHC-P |
Mouse
Rat
Human
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IP |
Human
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WB |
Mouse
Rat
Human
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All applications |
Sheep
Goat
Horse
Guinea pig
Cow
Cat
Dog
Pig
Non human primates
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Application | Abreviews | Notes |
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Flow Cyt |
Use at an assay dependent concentration.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
WB | (4) |
1/1000. Detects a band of approximately 64 kDa (predicted molecular weight: 64 kDa).
Milk recommended as blocking agent. |
IHC-P | (1) |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
ICC/IF | (2) |
1/1000.
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IP |
1/50.
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Notes |
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Flow Cyt
Use at an assay dependent concentration. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/1000. Detects a band of approximately 64 kDa (predicted molecular weight: 64 kDa). Milk recommended as blocking agent. |
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/1000. |
IP
1/50. |
Target
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Function
N6-methyltransferase that methylates adenosine residues of some mRNAs. N6-methyladenosine (m6A), which is present at internal sites of some mRNAs, may play a role in the efficiency of mRNA splicing, transport or translation. -
Tissue specificity
Widely expressed at low level. Expressed in spleen, thymus, prostate, testis, ovary, small intestine, colon and peripheral blood leukocytes. -
Sequence similarities
Belongs to the MT-A70-like family. -
Post-translational
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR. -
Cellular localization
Nucleus speckle. Colocalizes with speckles in interphase nuclei. Suggesting that it may be associated with nuclear pre-mRNA splicing components. - Information by UniProt
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Database links
- Entrez Gene: 56339 Human
- Entrez Gene: 56335 Mouse
- Entrez Gene: 361035 Rat
- Omim: 612472 Human
- SwissProt: Q86U44 Human
- SwissProt: Q8C3P7 Mouse
- Unigene: 168799 Human
- Unigene: 271759 Mouse
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Alternative names
- adoMet-binding subunit of the human mRNA (N6-adenosine)-methyltransferase antibody
- IME4 antibody
- M6A antibody
see all
Images
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Western blot - Anti-METTL3 antibody [EPR18810] (ab195352)This image is courtesy of an Abreview by Pedro Batista.All lanes : Anti-METTL3 antibody [EPR18810] (ab195352) at 1/1000 dilution
Lane 1 : Mouse embryonic stem cells
Lane 2 : Mouse embryonic stem cells (METTL3 Knock-out)
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat anti-Rabbit Polyclonal at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 64 kDa
Exposure time: 5 minutesBlocking step: 5% Milk for 30 minutes at 25oC.
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All lanes : Anti-METTL3 antibody [EPR18810] (ab195352) at 1/1000 dilution
Lane 1 : Mouse brain lysate prepared in 1%SDS Hot lysis method at 20 µg
Lane 2 : Mouse brain lysate prepared in RIPA lysis method at 20 mg/ml
Lane 3 : Mouse heart lysate prepared in 1%SDS Hot lysis method at 20 µg
Lane 4 : Mouse heart lysate prepared in RIPA lysis method at 20 mg/ml
Lane 5 : Mouse kidney lysate prepared in RIPA lysis method at 20 µg
Lanes 6-7 : Mouse spleen lysate prepared in spleen lysis method at 20 µg
Lane 8 : Rat brain lysate prepared in RIPA lysis method at 20 µg
Lane 9 : Rat kidney lysate prepared in RIPA lysis method at 20 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 64 kDa
Observed band size: 64 kDa -
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HCT116 (Human colorectal carcinoma cell line) cells labeling METTL3 with ab195352 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HCT116 cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with anti-alpha Tubulin mouse mAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (AlexaFluor®594) (ab150120) secondary antibody at 1/1000 dilution (red).
The negative controls are as follows:
1. ab195352 at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (AlexaFluor®594) (ab150120) secondary antibody at 1/1000 dilution.
2. anti-alpha Tubulin mouse mAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL3 antibody [EPR18810] (ab195352)
Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling METTL3 using ab195352 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Counterstain: Hematoxylin.
Inset image: negative control obtained using PBS instead of ab195352, and secondary antibody only.
Note: Nuclear staining on rat testis was observed.Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ab195352 staining METTL3 in the human cell line HEK-293 (Human epithelial cell line from embryonic kidney) by flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a 1/70 dilution, followed by Goat-Anti Rabbit IgG (Alexa Fluor® 488) secondary antibody at a 1/2000 dilution.
Isoytype control: Rabbit monoclonal IgG (Black)
Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)
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All lanes : Anti-METTL3 antibody [EPR18810] (ab195352) at 1/5000 dilution
Lane 1 : Raji (Human Burkitt's lymphoma cell line) whole cell lysate
Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 3 : HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate
Lane 4 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
Lane 5 : NCCIT (human pluripotent embryonal carcinoma) whole cell lysate
Lane 6 : F9 (Mouse embyro testicular cancer cell line) whole cell lysate
Lane 7 : Neuro-2a (Mouse neuroblastoma cells) whole cell lysate
Lane 8 : LLC1 (Mouse lung carcinoma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 64 kDa
Observed band size: 64 kDa
Exposure time: 10 seconds5% NFDM/TBST: Blocking and diluting buffer.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL3 antibody [EPR18810] (ab195352)
Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labeling METTL3 using ab195352 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Counterstain: Hematoxylin.
Inset image: negative control obtained using PBS instead of ab195352, and secondary antibody only.
Note: Nuclear staining on human bladder cancer was observed.Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling METTL3 with ab195352 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).
The negative controls are as follows:
1. ab195352 at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.
2. Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution. -
All lanes : Anti-METTL3 antibody [EPR18810] (ab195352) at 1/1000 dilution
Lane 1 : C6 (Rat glial tumor cells) whole cell lysate
Lane 2 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
Lane 4 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/100000 dilution
Predicted band size: 64 kDa
Observed band size: 64 kDa
Exposure time: 2 seconds5% NFDM/TBST: Blocking and diluting buffer.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL3 antibody [EPR18810] (ab195352)
Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling METTL3 using ab195352 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Counterstain: Hematoxylin.
Inset image: negative control obtained using PBS instead of ab195352, and secondary antibody only.
Note: Nuclear staining on mouse testis was observed.Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Anti-METTL3 antibody [EPR18810] (ab195352) at 1/5000 dilution + Human thymus lysate at 10 µg
Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 64 kDa
Observed band size: 64 kDa
Exposure time: 3 minutes5% NFDM/TBST: Blocking and diluting buffer.
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METTL3 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab195352 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab195352 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: Input: 10µg of HeLa whole cell lysate.
Lane 2: HeLa whole cell lysate following IP with ab195352.
Lane 3: Negative control: IP using Rabbit monoclonal IgG (ab172730) instead of ab195352 in HeLa whole cell lysates.
Blocking and dilution buffer and concentration: 5% NFDM/TBST. 1 second exposure.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (54)
ab195352 has been referenced in 54 publications.
- Pan X et al. METTL3 promotes adriamycin resistance in MCF-7 breast cancer cells by accelerating pri-microRNA-221-3p maturation in a m6A-dependent manner. Exp Mol Med N/A:N/A (2021). WB ; Human . PubMed: 33420414
- Han X et al. A suite of new Dre recombinase drivers markedly expands the ability to perform intersectional genetic targeting. Cell Stem Cell N/A:N/A (2021). IHC-Fr ; Mouse . PubMed: 33567267
- Xia TL et al. N(6)-methyladenosine-binding protein YTHDF1 suppresses EBV replication and promotes EBV RNA decay. EMBO Rep N/A:e50128 (2021). WB ; Human . PubMed: 33605073
- Han M et al. Abnormality of m6A mRNA Methylation Is Involved in Alzheimer's Disease. Front Neurosci 14:98 (2020). PubMed: 32184705
- Zhang C et al. METTL3 and N6-Methyladenosine Promote Homologous Recombination-Mediated Repair of DSBs by Modulating DNA-RNA Hybrid Accumulation. Mol Cell N/A:N/A (2020). WB, ICC/IF ; Human . PubMed: 32615088