Recombinant 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
Related conjugates and formulations
Overview
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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: 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.
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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.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18810 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
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 |
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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.
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ICC/IF | (2) |
1/1000.
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IP |
1/50.
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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 |
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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
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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
modificationsPhosphorylated 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
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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
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Alternative names
- adoMet-binding subunit of the human mRNA (N6-adenosine)-methyltransferase antibody
- IME4 antibody
- M6A antibody
see all
Images
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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 minutesBlocking step: 5% Milk for 30 minutes at 25oC.
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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 -
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. -
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. -
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. -
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. -
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. -
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.
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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. -
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.
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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)
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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 seconds5% NFDM/TBST: Blocking and diluting buffer.
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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.
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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 seconds5% NFDM/TBST: Blocking and diluting buffer.
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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.
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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 minutes5% NFDM/TBST: Blocking and diluting buffer.
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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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (237)
ab195352 has been referenced in 237 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
- 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
- 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
- Li Y et al. Single-cell m6A mapping in vivo using picoMeRIP-seq. Nat Biotechnol N/A:N/A (2023). WB ; Mouse . PubMed: 37349523