METTL3 Monoclonal antibody

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  WB analysis using 67733-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 67733-1-Ig (METTL3 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated Alpha Tubulin Monoclonal antibody (HRP-66031) as loading control.

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  WB analysis of A549 using 67733-1-Ig

  WB result of METTL3 antibody (67733-1-Ig; 1:10000; incubated at room temperature for 1.5 hours) with sh-Control and sh-METTL3 transfected A549 cells.

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  WB analysis of A549 using 67733-1-Ig

  WB result of METTL3 antibody (67733-1-Ig; 1:10000; incubated at room temperature for 1.5 hours) with sh-Control and sh-METTL3 transfected A549 cells.

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  IHC staining of human lung cancer using 67733-1-Ig

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 67733-1-Ig (METTL3 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human lung cancer using 67733-1-Ig

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 67733-1-Ig (METTL3 antibody) at dilution of 1:4000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human lung cancer using 67733-1-Ig

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 67733-1-Ig (METTL3 antibody) at dilution of 1:4000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of rat testis using 67733-1-Ig

  Immunohistochemical analysis of paraffin-embedded rat testis tissue slide using 67733-1-Ig (METTL3 antibody) at dilution of 1:4000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of rat testis using 67733-1-Ig

  Immunohistochemical analysis of paraffin-embedded rat testis tissue slide using 67733-1-Ig (METTL3 antibody) at dilution of 1:4000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human colon cancer using 67733-1-Ig

  Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 67733-1-Ig (METTL3 antibody) at dilution of 1:4000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human colon cancer using 67733-1-Ig

  Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 67733-1-Ig (METTL3 antibody) at dilution of 1:4000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human oesophagus cancer using 67733-1-Ig

  Immunohistochemical analysis of paraffin-embedded human oesophagus cancer tissue slide using 67733-1-Ig (METTL3 antibody) at dilution of 1:4000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human oesophagus cancer using 67733-1-Ig

  Immunohistochemical analysis of paraffin-embedded human oesophagus cancer tissue slide using 67733-1-Ig (METTL3 antibody) at dilution of 1:4000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human urothelial carcinoma using 67733-1-Ig

  Immunohistochemical analysis of paraffin-embedded human urothelial carcinoma tissue slide using 67733-1-Ig (METTL3 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human urothelial carcinoma using 67733-1-Ig

  Immunohistochemical analysis of paraffin-embedded human urothelial carcinoma tissue slide using 67733-1-Ig (METTL3 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human lung cancer using 67733-1-Ig

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 67733-1-Ig (METTL3 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of mouse testis using 67733-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed mouse testis tissue using METTL3 antibody (67733-1-Ig, Clone: 2D8H1 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

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  IF Staining of mouse testis using 67733-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed mouse testis tissue using METTL3 antibody (67733-1-Ig, Clone: 2D8H1 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

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  FC experiment of HEK-293 using 67733-1-Ig

  1X10^6 HEK-293 cells were intracellularly stained with 0.4 ug Anti-Human METTL3 (67733-1-Ig, Clone:2D8H1) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Control Antibody. Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

• Featured Product
• KD/KO Validated

METTL3 Monoclonal Antibody for WB, IHC, IF-P, FC (Intra), ELISA

Cat No. 67733-1-Ig

Clone No.2D8H1

Publications(28)

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse and More (1)

Applications

WB, IHC, IF-P, FC (Intra), IP, CoIP, RIP, ELISA

Synonyms

2D8H1, EC:2.1.1.348, hMETTL3, Methyltransferase-like protein 3, MT A70

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Tested Applications

• Positive WB detected in: HeLa cells, A549 cells, HEK-293 cells, HepG2 cells, Jurkat cells, K-562 cells
• Positive IHC detected in: human lung cancer tissue, human urothelial carcinoma tissue, human colon cancer tissue, human oesophagus cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF-P detected in: mouse testis tissue
• Positive FC (Intra) detected in: HEK-293 cells

This antibody is not recommended for immunocytofluorescent assays.

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:500-1:2000
• Immunofluorescence (IF)-P: IF-P : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 5 publications below
• WB: See 19 publications below
• IHC: See 5 publications below
• IF: See 8 publications below
• IP: See 4 publications below
• CoIP: See 1 publications below
• RIP: See 1 publications below

Product Information

67733-1-Ig targets METTL3 in WB, IHC, IF-P, FC (Intra), IP, CoIP, RIP, ELISA applications and shows reactivity with human, mouse samples.

• Tested Reactivity: human, mouse
• Cited Reactivity: human, mouse, rat
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen:

  CatNo: Ag7110

  Product name: Recombinant human METTL3 protein

  Source: e coli.-derived, T-HIS

  Tag: 6*His

  Domain: 229-580 aa of BC001650

  Sequence: LNQQSTKEQQSKKVSQEILELLNTTTAKEQSIVEKFRSRGRAQVQEFCDYGTKEECMKASDADRPCRKLHFRRIINKHTDESLGDCSFLNTCFHMDTCKYVHYEIDACMDSEAPGSKDHTPSQELALTQSVGGDSSADRLFPPQWICCDIRYLDVSILGKFAVVMADPPWDIHMELPYGTLTDDEMRRLNIPVLQDDGFLFLWVTGRAMELGRECLNLWGYERVDEIIWVKTNQLQRIIRTGRTGHWLNHGKEHCLVGVKGNPQGFNQGLDCDVIVAEVRSTSHKPDEIYGMIERLSPGTRKIELFGRPHNVQPNWITLGNQLDGIHLLDPDVVARFKQRYPDGIISKPKNL

• Full Name: methyltransferase like 3
• Calculated Molecular Weight: 64 kDa
• Observed Molecular Weight: 65-70 kDa
• GenBank Accession Number: BC001650
• Gene Symbol: METTL3
• Gene ID (NCBI): 56339
• RRID: AB_2918502
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• UNIPROT ID: Q86U44
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

METTL3 is a key S-adenosyl-L-methionine-binding subunit, which is component of a complex multicomponent enzyme that catalyzes the methylation of internal adenosine residues in eukaryotic mRNA, forming N6-methyladenosine. It contains 2 putative nuclear localization signals and 2 consensus methylation motifs. The calculated molecular weight of METTL3 is 64 kDa, but modified METTL3 is about 65-70 kDa.

Publications

• Human
• Mouse
• Rat
• Monkey
• Pig

Species	Application	Title
	WB,IHC	Adv Sci (Weinh); Elevated Histone Lactylation Mediates Ferroptosis Resistance in Endometriosis Through the METTL3-Regulated HIF1A/HMOX1 Signaling Pathway; Authors - Zongwen Liang
mouse	WB	Environ Pollut; RNA reading protein YTHDF2 mediates Benzo(k)fluoranthene induced male reproductive injury by regulating the stability of BCL2; Authors - Ya-Wen Li
mouse	WB	EMBO Rep; DDX5 inhibits inflammation by modulating m6A levels of TLR2/4 transcripts during bacterial infection; Authors - Jian Xu
human	IP	Cancer Lett; Therapeutic targeting m6A-guided miR-146a-5p signaling contributes to the melittin-induced selective suppression of bladder cancer.; Authors - Rucheng Yan
human	WB,IP,IF	J Invest Dermatol; NPTX2 Promotes EMT in Cutaneous Squamous Cell Carcinoma via Mettl3-Mediated m6A Methylation of Snail; Authors - Tong Kang
mouse,human	WB,IF	Am J Respir Cell Mol Biol; The Combined Effects of Circular RNA Methylation Promote Pulmonary Fibrosis.; Authors - Sha Wang ; KD Validated