METTL3 Recombinant Rabbit Monoclonal Antibody [JE58-20]
cat.: HA720002
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, IF-Cell
Clonality: Monoclonal
Clone number: JE58-20
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 64 kDa
Isotype: IgG
Immunogen: Recombinant protein within human METTL3 aa 1-300.
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.
Subcellular location: Nucleus, Nucleus speckle, Cytoplasm.
Recommended Dilutions:
  WB
  IHC-P
  IF-Cell
1:1,000
1:200-1:1,000
1:100
Uniprot #: SwissProt: Q86U44 Human | Q8C3P7 Mouse
Entrez Gene: 361035 Rat
Alternative names: adoMet-binding subunit of the human mRNA (N6-adenosine)-methyltransferase IME4 M6A Methyltransferase like protein 3 Methyltransferase-like protein 3 METTL3 mRNA (2'-O-methyladenosine-N(6)-)-methyltransferase mRNA m(6)A methyltransferase MT-A70 MTA70 MTA70_HUMAN N6 adenosine methyltransferase 70 kDa subunit N6-adenosine-methyltransferase 70 kDa subunit
Images
HA720002_1.jpg Fig1: 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.
HA720002_2.jpg Fig2: 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.
HA720002_3.jpg Fig3: 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.
HA720002_4.jpg Fig4: 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.
HA720002_5.jpg Fig5: 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.
HA720002_6.jpg Fig6: 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) 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.
HA720002_7.jpg Fig7: 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) 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.
HA720002_8.jpg Fig8: 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) 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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