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METTL3 Recombinant Rabbit Monoclonal Antibody [JE58-20]

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Specification

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Catalog# HA720002

METTL3 Recombinant Rabbit Monoclonal Antibody [JE58-20]

Application

  • WB

  • IHC-P

  • IF-Cell

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

METTL3 Recombinant Rabbit Monoclonal Antibody [JE58-20]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human METTL3 aa 1-300.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IF-Cell

Molecular Weight

Predicted band size: 64 kDa

Positive Control

Jurkat cell lysate, Neuro-2a cell lysate, PC-12 cell lysate, Human brain tissue lysate, Mouse brain tissue lysate, Rat brain tissue lysate, HeLa cell lysate, F9 cell lysate, Mouse heart tissue lysate, Rat heart tissue lysate, Mouse spleen tissue lysate, Rat spleen tissue lysate, HeLa, Neuro-2a, PC-12, human testis tissue, mouse testis tissue, rat testis tissue.

Conjugation

unconjugated

Clone Number

JE58-20

RRID

AB_3071943

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IHC-P

  • 1:200-1:1,000

  • IF-Cell

  • 1:100

Target

Function

This gene encodes the 70 kDa subunit of MT-A which is part of N6-adenosine-methyltransferase. This enzyme is involved in the posttranscriptional methylation of internal adenosine residues in eukaryotic mRNAs, forming N6-methyladenosine.

Background References

1. Li T. et. al. METTL3 facilitates tumor progression via an m(6)A-IGF2BP2-dependent mechanism in colorectal carcinoma. Mol Cancer. 2019 Jun

2. Choe J. et. al. mRNA circularization by METTL3-eIF3h enhances translation and promotes oncogenesis. Nature. 2018 Sep

Subcellular Location

Nucleus, Nucleus speckle, Cytoplasm.

UNIPROT

SwissProt: Q86U44 Human

SwissProt: Q8C3P7 Mouse

Entrez Gene: 361035 Rat

Synonyms

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

Expand

Images

  • Western blot analysis of METTL3 on different lysates with Rabbit anti-METTL3 antibody (<a href="/products/HA720002" style="font-weight: bold;text-decoration: underline;">HA720002</a>) at 1/1,000 dilution.<br /><br />Lane 1: Jurkat cell lysate<br />Lane 2: Neuro-2a cell lysate<br />Lane 3: PC-12 cell lysate<br />Lane 4: Human brain tissue lysate<br />Lane 5: Mouse brain tissue lysate<br />Lane 6: Rat brain tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 64 kDa<br />Observed band size: 72 kDa<br /><br />Exposure time: 26 seconds; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA720002" style="font-weight: bold;text-decoration: underline;">HA720002</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of METTL3 on different lysates with Rabbit anti-METTL3 antibody (HA720002) at 1/1,000 dilution.

    Lane 1: Jurkat cell lysate
    Lane 2: Neuro-2a cell lysate
    Lane 3: PC-12 cell lysate
    Lane 4: Human brain tissue lysate
    Lane 5: Mouse brain tissue lysate
    Lane 6: Rat brain tissue lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 64 kDa
    Observed band size: 72 kDa

    Exposure time: 26 seconds; ECL: K1801;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA720002) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of METTL3 on different lysates with Rabbit anti-METTL3 antibody (<a href="/products/HA720002" style="font-weight: bold;text-decoration: underline;">HA720002</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: F9 cell lysate (20 µg/Lane)<br />Lane 3: PC-12 cell lysate (20 µg/Lane)<br />Lane 4: Mouse heart tissue lysate (40 µg/Lane)<br />Lane 5: Rat heart tissue lysate (40 µg/Lane)<br />Lane 6: Mouse spleen tissue lysate (40 µg/Lane)<br />Lane 7: Rat spleen tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 64 kDa<br />Observed band size: 72 kDa<br /><br />Exposure time: 25 seconds; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA720002" style="font-weight: bold;text-decoration: underline;">HA720002</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of METTL3 on different lysates with Rabbit anti-METTL3 antibody (HA720002) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: F9 cell lysate (20 µg/Lane)
    Lane 3: PC-12 cell lysate (20 µg/Lane)
    Lane 4: Mouse heart tissue lysate (40 µg/Lane)
    Lane 5: Rat heart tissue lysate (40 µg/Lane)
    Lane 6: Mouse spleen tissue lysate (40 µg/Lane)
    Lane 7: Rat spleen tissue lysate (40 µg/Lane)

    Predicted band size: 64 kDa
    Observed band size: 72 kDa

    Exposure time: 25 seconds; ECL: K1801;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA720002) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HeLa cells labeling METTL3 with Rabbit anti-METTL3 antibody (<a href="/products/HA720002" style="font-weight: bold;text-decoration: underline;">HA720002</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-METTL3 antibody (<a href="/products/HA720002" style="font-weight: bold;text-decoration: underline;">HA720002</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling METTL3 with Rabbit anti-METTL3 antibody (HA720002) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-METTL3 antibody (HA720002) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunocytochemistry analysis of Neuro-2a cells labeling METTL3 with Rabbit anti-METTL3 antibody (<a href="/products/HA720002" style="font-weight: bold;text-decoration: underline;">HA720002</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-METTL3 antibody (<a href="/products/HA720002" style="font-weight: bold;text-decoration: underline;">HA720002</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of Neuro-2a cells labeling METTL3 with Rabbit anti-METTL3 antibody (HA720002) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-METTL3 antibody (HA720002) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunocytochemistry analysis of PC-12 cells labeling METTL3 with Rabbit anti-METTL3 antibody (<a href="/products/HA720002" style="font-weight: bold;text-decoration: underline;">HA720002</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-METTL3 antibody (<a href="/products/HA720002" style="font-weight: bold;text-decoration: underline;">HA720002</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of PC-12 cells labeling METTL3 with Rabbit anti-METTL3 antibody (HA720002) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-METTL3 antibody (HA720002) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-METTL3 antibody (<a href="/products/HA720002" style="font-weight: bold;text-decoration: underline;">HA720002</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA720002" style="font-weight: bold;text-decoration: underline;">HA720002</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-METTL3 antibody (HA720002) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA720002) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-METTL3 antibody (<a href="/products/HA720002" style="font-weight: bold;text-decoration: underline;">HA720002</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA720002" style="font-weight: bold;text-decoration: underline;">HA720002</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-METTL3 antibody (HA720002) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA720002) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-METTL3 antibody (<a href="/products/HA720002" style="font-weight: bold;text-decoration: underline;">HA720002</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA720002" style="font-weight: bold;text-decoration: underline;">HA720002</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-METTL3 antibody (HA720002) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA720002) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

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