Recombinant
RabMAb

Recombinant Anti-METTL3 antibody [EPR18810] - BSA and Azide free (ab221795)

Rabbit recombinant monoclonal METTL3 antibody [EPR18810]. Validated in WB, IP, IHC, Flow Cyt, ICC/IF and tested in Mouse, Rat, Human. Immunogen corresponding to recombinant fragment.

Overview

  • Product name

    Anti-METTL3 antibody [EPR18810] - BSA and Azide free
    See all METTL3 primary antibodies
  • Description

    Rabbit monoclonal [EPR18810] to METTL3 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, Flow Cyt, IP, IHC-Fr, ICC/IF, WBmore 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 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

    Ab221795 is the carrier-free version of ab195352. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab221795 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab221795 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
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 64 kDa (predicted molecular weight: 64 kDa).

Milk recommended as blocking agent.

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

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195352).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195352).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195352).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195352).

References

ab221795 has not yet been referenced specifically in any publications.

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