Revision 7

#86132Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, W-S, IP, IHC-P, ChIP

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

70

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q86U44

Entrez-Gene Id:

56339

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunoprecipitation 1:50
Immunohistochemistry (Paraffin) 1:200 - 1:800
Chromatin IP 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

METTL3 (E3F2A) Rabbit mAb recognizes endogenous levels of total METTL3 protein.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human METTL3 protein.

Background

Methyltransferase-like protein 3 (METTL3) and methyltransferase-like protein 14 (METTL14) are the two catalytic subunits of an N6-methyltransferase complex that methylates adenosine residues in RNA (1). Methylation of adenosine residues regulates mRNA splicing, processing, translation efficiency, editing and stability, in addition to regulating primary miRNA processing, and is critical for proper regulation of the circadian clock, embryonic stem cell self-renewal, immune tolerance, response to various stimuli, meiosis and mouse fertility (2,3). In this complex, METTL3 functions as the catalytic methyltransferase subunit and METTL14 functions as the target recognition subunit by binding to RNA (4). In addition, the Wilms' tumor 1-associating protein (WTAP) functions as a regulatory subunit and is required for accumulation of the complex to nuclear speckles, which are sites of RNA processing (5). Several studies suggest a role for this complex in cancer. METTL3 expression is elevated in lung adenocarcinoma where it promotes growth, survival, and invasion of human lung cancer cells (6). In addition, WTAP is overexpressed in a number of different cancers and positively regulates cell migration and invasion in glioblastoma and cholangiocarcinoma (7,8).

  1. Liu, J. et al. (2014) Nat Chem Biol 10, 93-5.
  2. Yue, Y. et al. (2015) Genes Dev 29, 1343-55.
  3. Wu, R. et al. (2016) Mol Biotechnol 58, 450-9.
  4. Wang, X. et al. (2016) Nature 534, 575-8.
  5. Ping, X.L. et al. (2014) Cell Res 24, 177-89.
  6. Lin, S. et al. (2016) Mol Cell 62, 335-45.
  7. Jin, D.I. et al. (2012) Cancer Sci 103, 2102-9.
  8. Jo, H.J. et al. (2013) J Gastroenterol 48, 1271-82.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) ChIP: Chromatin IP

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

使用に関する制限

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Revision 7
#86132

METTL3 (E3F2A) Rabbit mAb

Western Blotting Image 1: METTL3 (E3F2A) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using METTL3 (E3F2A) Rabbit mAb.
Western Blotting Image 1: METTL3 (E3F2A) Rabbit mAb Expand Image
Simple Western™ analysis of lysates (0.1 mg/mL) from COS-7 cells using METTL3 (E3F2A) Rabbit mAb #86132. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Immunoprecipitation Image 1: METTL3 (E3F2A) Rabbit mAb Expand Image
Immunoprecipitation of METTL3 from 293T cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is METTL3 (E3F2A) Rabbit mAb. Western blot analysis was performed using METTL3 (E3F2A) Rabbit mAb and Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127.
Immunohistochemistry Image 1: METTL3 (E3F2A) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using METTL3 (E3F2A) Rabbit mAb.
Immunohistochemistry Image 2: METTL3 (E3F2A) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using METTL3 (E3F2A) Rabbit mAb.
Immunohistochemistry Image 3: METTL3 (E3F2A) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human large cell neuroendocrine lung carcinoma using METTL3 (E3F2A) Rabbit mAb.
Immunohistochemistry Image 4: METTL3 (E3F2A) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human B-cell non-Hodgkin lymphoma using METTL3 (E3F2A) Rabbit mAb.
Immunohistochemistry Image 5: METTL3 (E3F2A) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal human kidney using METTL3 (E3F2A) Rabbit mAb.
Immunohistochemistry Image 6: METTL3 (E3F2A) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal human pancreas using METTL3 (E3F2A) Rabbit mAb.
Immunohistochemistry Image 7: METTL3 (E3F2A) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal human heart using METTL3 (E3F2A) Rabbit mAb.
Immunohistochemistry Image 8: METTL3 (E3F2A) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse brain using METTL3 (E3F2A) Rabbit mAb.
Immunohistochemistry Image 9: METTL3 (E3F2A) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse testis using METTL3 (E3F2A) Rabbit mAb.
Immunohistochemistry Image 10: METTL3 (E3F2A) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human ovarian adenocarcinoma using METTL3 (E3F2A) Rabbit mAb (left) or a METTL3 Rabbit mAb (right). These two antibodies detect unique, non-overlapping epitopes on METTL3. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining.
Immunohistochemistry Image 11: METTL3 (E3F2A) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using METTL3 (E3F2A) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right). 
Chromatin Immunoprecipitation Image 1: METTL3 (E3F2A) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from mES cells and either METTL3 (E3F2A) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using SimpleChIP® Mouse Sox2 Exon1 Primers #30180, mouse KLF2 intron 2 primers, and SimpleChIP® Mouse MYT-1 Promoter Primers #8985. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.