Name: Anti-METTL3 antibody [EPR18810]
Brand: RabMAb
SKU: ab195352
Rating: 5 (8 reviews)

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Primary

Secondary

This product Rabbit Anti-METTL3 antibody [EPR18810] (ab195352)
Flow Cyt, IHC-Fr, WB, IHC-P, ICC/IF, IP

Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
ICC/IF, Flow Cyt, IHC-P, ELISA, IHC-Fr

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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

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

Run BLAST with Run BLAST with
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

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.

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

Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18810
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human METTL3 protein (ab163377)
Related Products
- Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)
- VeriBlot for IP Detection Reagent (HRP) (ab131366)

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.

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.
Target information above from: UniProt accession Q86U44 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
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
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

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 minutes

Blocking step: 5% Milk for 30 minutes at 25^oC.
See Abreview
Western blot - Anti-METTL3 antibody [EPR18810] (ab195352)

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.
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.
Immunocytochemistry/ Immunofluorescence - Anti-METTL3 antibody [EPR18810] (ab195352)

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.
Flow Cytometry - Anti-METTL3 antibody [EPR18810] (ab195352)

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)
Western blot - Anti-METTL3 antibody [EPR18810] (ab195352)

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.
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.
Immunocytochemistry/ Immunofluorescence - Anti-METTL3 antibody [EPR18810] (ab195352)

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.
Western blot - Anti-METTL3 antibody [EPR18810] (ab195352)

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.
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.
Western blot - Anti-METTL3 antibody [EPR18810] (ab195352)

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.
Western blot - Anti-METTL3 antibody [EPR18810] (ab195352)

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.
Immunoprecipitation - Anti-METTL3 antibody [EPR18810] (ab195352)

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.
Immunohistochemistry (Frozen sections) - Anti-METTL3 antibody [EPR18810] (ab195352)This image is courtesy of an abreview submitted by Bryan Niedenberger

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

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Datasheet

This product has been referenced in:

Nettersheim D et al. N6-Methyladenosine detected in RNA of testicular germ cell tumors is controlled by METTL3, ALKBH5, YTHDC1/F1/F2, and HNRNPC as writers, erasers, and readers. Andrology N/A:N/A (2019). WB ; Human . Read more (PubMed: 30903744) »
Zhang J et al. Excessive miR-25-3p maturation via N6-methyladenosine stimulated by cigarette smoke promotes pancreatic cancer progression. Nat Commun 10:1858 (2019). WB ; Human . Read more (PubMed: 31015415) »

See all 12 Publications for this product

Publishing research using ab195352? Please let us know so that we can cite the reference in this datasheet.

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1-8 of 8 Abreviews or Q&A

Application

Western blot

Sample

Human Cell lysate - whole cell (HEK293)

Gel Running Conditions

Reduced Denaturing (4-12% Bis-tris)

Loading amount

20 µg

Specification

HEK293

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 Mar 22 2019

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

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

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

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

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

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

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

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Recombinant Anti-METTL3 antibody [EPR18810] (ab195352)

Overview

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Images

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Datasheets and documents

References

This product has been referenced in:

Customer reviews and Q&As

Western blot abreview for Anti-METTL3 antibody [EPR18810]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-METTL3 antibody [EPR18810]

Immunocytochemistry/ Immunofluorescence abreview for Anti-METTL3 antibody [EPR18810]

Western blot abreview for Anti-METTL3 antibody [EPR18810]

Western blot abreview for Anti-METTL3 antibody [EPR18810]

Immunohistochemistry (Frozen sections) abreview for Anti-METTL3 antibody [EPR18810]

Immunocytochemistry/ Immunofluorescence abreview for Anti-Methyltransferase like 3 antibody [EPR18810]

Western blot abreview for Anti-Methyltransferase like 3 antibody [EPR18810]