METTL14 Recombinant Rabbit Monoclonal Antibody [PSH07-85]
cat.: HA722910
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat, Monkey
Applications: WB, IF-Cell, IHC-P, FC, IP
Clonality: Monoclonal
Clone number: PSH07-85
Form: Liquid
Storage condition: 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.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 52 kDa
Isotype: IgG
Immunogen: Recombinant protein within human METTL14 aa 1-150.
Positive control: A431 cell lysate, Raji cell lysate, PC-12 cell lysate, COS-1 cell lysate, Mouse testis tissue lysate, Rat testis tissue lysate, A431, human kidney tissue, human lung tissue, mouse testis tissue, rat kidney tissue, rat lung tissue, rat testis tissue.
Subcellular location: Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
  IP
1:5,000
1:100
1:200-1:1,000
1:1,000
1-2μg/sample
Uniprot #: SwissProt: Q9HCE5 Human | Q3UIK4 Mouse
Entrez Gene: 295428 Rat
Alternative names: KIAA1627 MET14_HUMAN Methyltransferase like 14 Methyltransferase-like protein 14 METTL14 N6-adenosine-methyltransferase subunit METTL14 OTTHUMP00000219837
Images
HA722910_1.jpg Fig1: Western blot analysis of METTL14 on different lysates with Rabbit anti-METTL14 antibody (HA722910) 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)

Predicted band size: 52 kDa
Observed band size: 60 kDa

Exposure time: 30 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 (HA722910) at 1/5,000 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.
HA722910_2.jpg Fig2: Immunocytochemistry analysis of A431 cells labeling METTL14 with Rabbit anti-METTL14 antibody (HA722910) 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 (HA722910) 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.
HA722910_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-METTL14 antibody (HA722910) 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 (HA722910) 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.
HA722910_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human lung tissue with Rabbit anti-METTL14 antibody (HA722910) 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 (HA722910) 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.
HA722910_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-METTL14 antibody (HA722910) 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 (HA722910) 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.
HA722910_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-METTL14 antibody (HA722910) 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 (HA722910) 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.
HA722910_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded rat lung tissue with Rabbit anti-METTL14 antibody (HA722910) 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 (HA722910) 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.
HA722910_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-METTL14 antibody (HA722910) 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 (HA722910) 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.
HA722910_9.jpg Fig9: Flow cytometric analysis of A431 cells labeling METTL14.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA722910, 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 (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
HA722910_10.jpg Fig10: METTL14 was immunoprecipitated from 0.2 mg A431 cell lysate with HA722910 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using HA722910 at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: A431 cell lysate (input)
Lane 2: HA722910 IP in A431 cell lysate
Lane 3: Rabbit IgG instead of HA722910 in A431 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 1 minute; ECL: K1801
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.