Overview

  • Product name
    Anti-METTL3 antibody [EPR18810]
    See all METTL3 primary antibodies
  • Description
    Rabbit monoclonal [EPR18810] to METTL3
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, IHC-Fr, WB, IHC-P, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Pig
    Predicted to work with: Sheep, Goat, Horse, Guinea pig, Cow, Cat, Dog, Non human primates
  • Immunogen

    Recombinant fragment within Human METTL3 aa 1-250. The exact sequence is proprietary.
    Database link: Q86U44

  • 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, heart and kidney 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.
  • General notes

    Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).

    See other anti-rabbit secondary antibodies that can be used with this antibody.

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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.

Application Abreviews Notes
Flow Cyt Use at an assay dependent concentration.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-Fr 1/1000.
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

  • 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
    modifications
    Phosphorylated 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
  • Database links
  • Alternative names
    • adoMet-binding subunit of the human mRNA (N6-adenosine)-methyltransferase antibody
    • IME4 antibody
    • M6A antibody
    • Methyltransferase like protein 3 antibody
    • Methyltransferase-like protein 3 antibody
    • METTL3 antibody
    • mRNA (2'-O-methyladenosine-N(6)-)-methyltransferase antibody
    • mRNA m(6)A methyltransferase antibody
    • MT-A70 antibody
    • MTA70 antibody
    • MTA70_HUMAN antibody
    • N6 adenosine methyltransferase 70 kDa subunit antibody
    • N6-adenosine-methyltransferase 70 kDa subunit antibody
    see all

Images

  • 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 minutes


    Blocking step: 5% Milk for 30 minutes at 25oC.

    See Abreview

  • 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 seconds


    5% NFDM/TBST: Blocking and diluting buffer.

  • 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.

  • 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.

  • 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)

  • All lanes : Anti-METTL3 antibody [EPR18810] (ab195352) at 1/1000 dilution

    Lane 1 : mouse brain lysate
    Lane 2 : mouse heart lysate
    Lane 3 : mouse kidney 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: 30 seconds


    5% NFDM/TBST: Blocking and diluting buffer.

  • 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.

  • 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 seconds


    5% NFDM/TBST: Blocking and diluting buffer.

  • 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.

  • 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 minutes


    5% NFDM/TBST: Blocking and diluting buffer.

  • Anti-METTL3 antibody [EPR18810] (ab195352) at 1/1000 dilution + rat brain lysate at 10 µg

    Secondary
    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: 20 seconds


    5% NFDM/TBST: Blocking and diluting buffer.

  • 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 secondary antibody (HRP) (ab131366), was used as secondary antibody 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.

  • Immunohistochemical analysis of adult mouse frozen testis sections labelling METTL3 with ab195352 at dilution of 1/1000. The secondary antibody used was ab150081at a dilution of 1/500. The section was fixed with Paraformaldehyde and permeabilised with 0.1% Triton X-100 in PBS. The sample was counterstained with Phalloidin (red) which labels F-actin.

    See Abreview

References

This product has been referenced in:
  • Wen J  et al. Zc3h13 Regulates Nuclear RNA m6A Methylation and Mouse Embryonic Stem Cell Self-Renewal. Mol Cell 69:1028-1038.e6 (2018). WB, ICC/IF ; Mouse . Read more (PubMed: 29547716) »
  • Song T  et al. Maternal obesity aggravates the abnormality of porcine placenta by increasing N6-methyladenosine. Int J Obes (Lond) N/A:N/A (2018). WB ; Pig . Read more (PubMed: 29795472) »
See all 7 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (bladder carcinoma)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA, PH9
Permeabilization
No
Specification
bladder carcinoma
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Apr 27 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Pig Cell (Swine Testicle cells (ST))
Permeabilization
Yes - 0.1% triton X-100
Specification
Swine Testicle cells (ST)
Fixative
Paraformaldehyde

Mr. Robert Jansens

Verified customer

Submitted Oct 27 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse embryonic stem cells)
Gel Running Conditions
Reduced Denaturing (4-12%)
Loading amount
15 µg
Treatment
wild type and METTL3 knock-out
Specification
mouse embryonic stem cells
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Pedro Batista

Verified customer

Submitted Oct 29 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa)
Gel Running Conditions
Non-reduced Denaturing (4-12% gel)
Loading amount
10 µg
Specification
HeLa
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Sep 07 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Testis, adult)
Permeabilization
Yes - 0.1% Triton X-100 in PBS
Specification
Testis, adult
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C
Fixative
Paraformaldehyde

Mr. Bryan Niedenberger

Verified customer

Submitted Sep 02 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (human embryonic stem cells)
Permeabilization
Yes - 0.25% TritonX-100
Specification
human embryonic stem cells
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 37°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jan 21 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (human embryonic stem cells)
Gel Running Conditions
Reduced Denaturing (4-20%)
Loading amount
30 µg
Specification
human embryonic stem cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C

Abcam user community

Verified customer

Submitted Jan 21 2016

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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