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

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Western blot - 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: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)
Knockout validated
Reacts with: Mouse, Rat, Human

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

Product name

Anti-METTL3 antibody [EPR18810]
See all METTL3 primary antibodies
Description

Rabbit monoclonal [EPR18810] to METTL3
Host species

Rabbit
Tested applications

Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)more details
Species reactivity

Reacts with: Mouse, Rat, Human
Predicted to work with: Sheep, Goat, Horse, Guinea pig, Cow, Cat, Dog, Pig, Non human primates
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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, mouse skin, rat skin. ICC/IF: HCT116 and HeLa, NIH/3T3 cells. IP: HeLa cell lysate.
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
For more information see here.
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.

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18810
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
- 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)

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.

Application	Abreviews	Notes
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.
IP		1/50.
Flow Cyt (Intra)		Use at an assay dependent concentration. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Notes
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.
Flow Cyt (Intra) Use at an assay dependent concentration. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

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 1/1000 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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL3 antibody [EPR18810] (ab195352)

Immunohistochemical analysis of paraffin-embedded Rat skin tissue labeling METTL3 using ab195352 at 1/2000 dilution, followed by a Ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Counterstain: Hematoxylin. Nuclear staining on Rat skin. The section was incubated with ab195352 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Perform Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes.

Inset image: negative control - secondary antibody only.
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 1/2000 dilution, followed by a Ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Counterstain: Hematoxylin. Nuclear staining on Rat testis. The section was incubated with ab195352 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Perform Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes.

Inset image: negative control - secondary antibody only.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL3 antibody [EPR18810] (ab195352)

Immunohistochemical analysis of paraffin-embedded Mouse skin labeling METTL3 using ab195352 at 1/2000 dilution, followed by a Ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Counterstain: Hematoxylin. Nuclear staining on Mouse skin. The section was incubated with ab195352 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Perform Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes.

Inset image: negative control - secondary antibody only.
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 1/2000 dilution, followed by a Ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Counterstain: Hematoxylin. Nuclear staining on Mouse testis. The section was incubated with ab195352 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Perform Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes.

Inset image: negative control - secondary antibody only.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL3 antibody [EPR18810] (ab195352)

Immunohistochemical analysis of paraffin-embedded Human ovarian cancer tissue labeling METTL3 using ab195352 at 1/2000 dilution, followed by a Ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Counterstain: Hematoxylin. Nuclear staining on human ovarian cancer. The section was incubated with ab195352 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Perform Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes.

Inset image: negative control - secondary antibody only.
Immunocytochemistry/ Immunofluorescence - Anti-METTL3 antibody [EPR18810] (ab195352)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labeling METTL3 with ab195352 at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150081) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in NIH/3T3 cells. The nuclear counter stain is DAPI (blue). Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) ab195889 was used at 1/200 dilution as the counterstain.
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.
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.
Flow Cytometry (Intracellular) - Anti-METTL3 antibody [EPR18810] (ab195352)

ab195352 staining METTL3 in the human cell line HEK-293 (Human epithelial cell line from embryonic kidney) by intracellular 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 antibodyat a1/2000 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 1/5000 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 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.
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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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.
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.
Anti-METTL3 antibody [EPR18810] (ab195352)

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

Click here to view the general protocols

SDS download

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Language
Datasheet download

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

ab195352 has been referenced in 152 publications.

Kong H et al. Long intergenic non-protein coding RNA 1273 confers sorafenib resistance in hepatocellular carcinoma via regulation of methyltransferase 3. Bioengineered 13:3108-3121 (2022). WB ; Human . PubMed: 35037556
Han D et al. Dynamic assembly of the mRNA m6A methyltransferase complex is regulated by METTL3 phase separation. PLoS Biol 20:e3001535 (2022). ICC ; Human . PubMed: 35143475
Su R et al. METTL16 exerts an m6A-independent function to facilitate translation and tumorigenesis. Nat Cell Biol 24:205-216 (2022). ICC ; Human . PubMed: 35145225
Wei K et al. Methylation recognition protein YTH N6-methyladenosine RNA binding protein 1 (YTHDF1) regulates the proliferation, migration and invasion of osteosarcoma by regulating m6A level of CCR4-NOT transcription complex subunit 7 (CNOT7). Bioengineered 13:5236-5250 (2022). IP ; Human . PubMed: 35156522
Zhang L et al. METTL3 suppresses neuropathic pain via modulating N6-methyladenosine-dependent primary miR-150 processing. Cell Death Discov 8:80 (2022). WB, IP, IF ; Rat . PubMed: 35210391

View all Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Application

Immunocytochemistry/ Immunofluorescence

Sample

Pig Cell (Swine Testicle cells (ST))

Permeabilization

Yes - 0.1% triton X-100

Specification

Swine Testicle cells (ST)

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Mr. Robert Jansens

Verified customer

Submitted Oct 27 2017

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

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jan 21 2016

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (152)

Customer reviews and Q&As

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

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

Post-translational
modifications