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    products/primary-antibodies/mettl3-antibody-epr18810-ab195352.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-METTL3 antibody [EPR18810] (ab195352)

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  • Certificate of Compliance
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Western blot - Anti-METTL3 antibody [EPR18810] (ab195352)
  • Western blot - Anti-METTL3 antibody [EPR18810] (ab195352)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL3 antibody [EPR18810] (ab195352)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL3 antibody [EPR18810] (ab195352)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL3 antibody [EPR18810] (ab195352)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL3 antibody [EPR18810] (ab195352)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL3 antibody [EPR18810] (ab195352)
  • Immunocytochemistry/ Immunofluorescence - Anti-METTL3 antibody [EPR18810] (ab195352)
  • Immunocytochemistry/ Immunofluorescence - Anti-METTL3 antibody [EPR18810] (ab195352)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL3 antibody [EPR18810] (ab195352)
  • Flow Cytometry (Intracellular) - Anti-METTL3 antibody [EPR18810] (ab195352)
  • Western blot - Anti-METTL3 antibody [EPR18810] (ab195352)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL3 antibody [EPR18810] (ab195352)
  • Immunocytochemistry/ Immunofluorescence - Anti-METTL3 antibody [EPR18810] (ab195352)
  • Western blot - Anti-METTL3 antibody [EPR18810] (ab195352)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL3 antibody [EPR18810] (ab195352)
  • Western blot - Anti-METTL3 antibody [EPR18810] (ab195352)
  • Immunoprecipitation - Anti-METTL3 antibody [EPR18810] (ab195352)
  • 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

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

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View more associated products

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: 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.

Properties

  • 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

    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA / Nucleotides
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • RNA Processing
    • Other

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-METTL3 antibody [EPR18810] (ab216936)
    • Alexa Fluor® 647 Anti-METTL3 antibody [EPR18810] (ab217109)
    • Anti-METTL3 antibody [EPR18810] - BSA and Azide free (ab221795)
  • 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)

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.

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.

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.
  • 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

    Images

    • Western blot - Anti-METTL3 antibody [EPR18810] (ab195352)
      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 25oC.

      See Abreview

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

    Protocols

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

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (165)

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

    ab195352 has been referenced in 165 publications.

    • Fu H  et al. m6A contributes to a pro-survival state in GC-2 cells by facilitating DNA damage repair: Novel perspectives on the mechanism underlying DEHP genotoxicity in male germ cells. Sci Total Environ 859:160432 (2023). WB, ICC ; Mouse . PubMed: 36423848
    • Zhao T  et al. N6-methyladenosine plays a dual role in arsenic carcinogenesis by temporal-specific control of core target AKT1. J Hazard Mater 445:130468 (2023). RIP ; Human . PubMed: 36444808
    • Dai M  et al. BPDE, the Migration and Invasion of Human Trophoblast Cells, and Occurrence of Miscarriage in Humans: Roles of a Novel lncRNA-HZ09. Environ Health Perspect 131:17009 (2023). WB ; Human . PubMed: 36719213
    • Li J  et al. Integrated analysis of RNA methylation regulators crosstalk and immune infiltration for predictive and personalized therapy of diabetic nephropathy. Hum Genomics 17:6 (2023). ICC ; Human . PubMed: 36765416
    • Zhang J  et al. TWIK-related acid-sensitive K+ channel 2 promotes renal fibrosis by inducing cell-cycle arrest. iScience 25:105620 (2022). WB ; Mouse . PubMed: 36465115
    View all Publications for this product

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    1-8 of 8 Abreviews

    Western blot abreview for Anti-METTL3 antibody [EPR18810]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    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 Mar 22 2019

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

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    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 Apr 27 2018

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

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

    Western blot abreview for Anti-METTL3 antibody [EPR18810]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    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.

    Pedro Batista

    Verified customer

    Submitted Oct 29 2016

    Western blot abreview for Anti-METTL3 antibody [EPR18810]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    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 Sep 07 2016

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    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. Bryan Niedenberger

    Verified customer

    Submitted Sep 02 2016

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    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

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    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

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