WB
Western blot analysis of METTL14 on different lysates with Rabbit anti-METTL14 antibody at 1/5,000 dilution. Lane 1: A431 cell lysate (20 µg/Lane) Lane 2: Raji cell lysate (20 µg/Lane) Lane 3: PC-12 cell lysate (20 µg/Lane) Lane 4: COS-1 cell lysate (20 µg/Lane) Lane 5: Mouse testis tissue lysate (40 µg/Lane) Lane 6: Rat testis tissue lysate (40 µg/Lane) Exposure time: 30 seconds; 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 at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-METTL14 antibody 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 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.ICC/IF
Immunocytochemistry analysis of A431 cells labeling METTL14 with Rabbit anti-METTL14 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-METTL14 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of A431 cells labeling METTL14. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
METTL14 was immunoprecipitated from 0.2 mg A431 cell lysate with Rabbit anti-METTL14 antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate usingRabbit anti-METTL14 antibody at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: A431 cell lysate (input) Lane 2: Rabbit anti-METTL14 antibody IP in A431 cell lysate Lane 3: Rabbit IgG instead of Rabbit anti-METTL14 antibody in A431 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 1 minute.| Product Name | METTL14 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within human METTL14 aa 1-150. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCIPWB |
| WB:1:5000 IHC:1:200-1:1000 ICC:1:100 FC:1:1000 IP:1-2μg/sample |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | METTL14 |
|---|---|
| Gene Synonyms | hMETTL14 |
| Gene Full Name | methyltransferase 14, N6-adenosine-methyltransferase non-catalytic subunit |
| Gene Summary | Enables mRNA binding activity and mRNA m(6)A methyltransferase activity. Involved in mRNA modification; mRNA splicing, via spliceosome; and mRNA stabilization. Located in nucleoplasm. Is active in RNA N6-methyladenosine methyltransferase complex. [provided by Alliance of Genome Resources, Nov 2024] |
| Alternative Names | KIAA1627 antibody MET14_HUMAN antibody Methyltransferase like 14 antibody Methyltransferase-like protein 14 antibody METTL14 antibody N6-adenosine-methyltransferase subunit METTL14 antibody OTTHUMP00000219837 antibody |
| Molecular Weight(MW) | 52kDa(Observed band size: 60kDa) |
| Cellular Localization | Nucleus. |
WB
Western blot analysis of METTL14 on different lysates with Rabbit anti-METTL14 antibody at 1/5,000 dilution. Lane 1: A431 cell lysate (20 µg/Lane) Lane 2: Raji cell lysate (20 µg/Lane) Lane 3: PC-12 cell lysate (20 µg/Lane) Lane 4: COS-1 cell lysate (20 µg/Lane) Lane 5: Mouse testis tissue lysate (40 µg/Lane) Lane 6: Rat testis tissue lysate (40 µg/Lane) Exposure time: 30 seconds; 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 at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-METTL14 antibody 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 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.
ICC/IF
Immunocytochemistry analysis of A431 cells labeling METTL14 with Rabbit anti-METTL14 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-METTL14 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of A431 cells labeling METTL14. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
METTL14 was immunoprecipitated from 0.2 mg A431 cell lysate with Rabbit anti-METTL14 antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate usingRabbit anti-METTL14 antibody at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: A431 cell lysate (input) Lane 2: Rabbit anti-METTL14 antibody IP in A431 cell lysate Lane 3: Rabbit IgG instead of Rabbit anti-METTL14 antibody in A431 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 1 minute.| Application Notes | WB:1:5000 IHC:1:200-1:1000 ICC:1:100 FC:1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
