Product Name
VIRMA Recombinant Rabbit Monoclonal Antibody [PSH08-28] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within human VIRMA aa 1,431-1,730.
Validated Applications
WB, IF-Cell, IHC-P, FC, IP
Target Molecular Weight
Predicted band size: 202 kDa
Positive Control
K-562 cell lysate, MCF7 cell lysate, HeLa cell lysate, PC-3 cell lysate, T-47D cell lysate, SK-OV-3 cell lysate, COS-1 cell lysate, PC-12 cell lysate, C6 cell lysate, HeLa, PC-12, human colon cancer tissue, human stomach tissue, human testis tissue, rat stomach tissue, rat testis tissue.
Storage Instructions
Store at 2-8℃. Avoid freeze.
Purification Method
Protein A affinity purified.
Function
Associated component of the WMM complex, a complex that mediates N6-methyladenosine (m6A) methylation of RNAs, a modification that plays a role in the efficiency of mRNA splicing and RNA processing. Acts as a key regulator of m6A methylation by promoting m6A methylation of mRNAs in the 3'-UTR near the stop codon: recruits the catalytic core components METTL3 and METTL14, thereby guiding m6A methylation at specific sites. Required for mRNA polyadenylation via its role in selective m6A methylation: m6A methylation of mRNAs in the 3'-UTR near the stop codon correlating with alternative polyadenylation (APA).
Background References
1. Li N et al. KIAA1429/VIRMA promotes breast cancer progression by m(6) A-dependent cytosolic HAS2 stabilization. EMBO Rep. 2023 Oct
2. Xu H et al. VIRMA facilitates intrahepatic cholangiocarcinoma progression through epigenetic augmentation of TMED2 and PARD3B mRNA stabilization. J Gastroenterol. 2023 Sep
Subcellular Location
Nucleus speckle, Nucleus, nucleoplasm, Cytoplasm.
Synonyms
DKFZp434I116 antibody
DKFZp781B2117 antibody
fSAP121 antibody
KIAA1429 antibody
MGC138493 antibody
MGC141940 antibody
MSTP054 antibody
Protein virilizer homolog antibody
VIR_HUMAN antibody
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This data was developed using HA722972, the same antibody clone in a different buffer formulation.
Western blot analysis of VIRMA on different lysates with Rabbit anti-VIRMA antibody (HA722972) at 1/2,000 dilution.
Lane 1: K-562 cell lysate
Lane 2: MCF7 cell lysate
Lane 3: HeLa cell lysate
Lane 4: PC-3 cell lysate
Lane 5: T-47D cell lysate
Lane 6: SK-OV-3 cell lysate
Lane 7: COS-1 cell lysate
Lane 8: PC-12 cell lysate
Lane 9: C6 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 202 kDa
Observed band size: 240 kDa
Exposure time: 10 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 (HA722972) at 1/2,000 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.
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This data was developed using HA722972, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of HeLa cells labeling VIRMA with Rabbit anti-VIRMA antibody (HA722972) 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-VIRMA antibody (HA722972) 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 (HA601187, 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.
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This data was developed using HA722972, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of PC-12 cells labeling VIRMA with Rabbit anti-VIRMA antibody (HA722972) at 1/50 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-VIRMA antibody (HA722972) at 1/50 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 (HA601187, 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.
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This data was developed using HA722972, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-VIRMA antibody (HA722972) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA722972) 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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This data was developed using HA722972, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human stomach tissue with Rabbit anti-VIRMA antibody (HA722972) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA722972) 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.
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This data was developed using HA722972, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-VIRMA antibody (HA722972) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA722972) 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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This data was developed using HA722972, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat stomach tissue with Rabbit anti-VIRMA antibody (HA722972) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA722972) 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.
-
This data was developed using HA722972, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-VIRMA antibody (HA722972) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA722972) 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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This data was developed using HA722972, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of HeLa cells labeling VIRMA.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA722972, 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).
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This data was developed using HA722972, the same antibody clone in a different buffer formulation.
VIRMA was immunoprecipitated from 0.2 mg K-562 cell lysate with HA722972 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA722972 at 1/1,000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody (M1208-2) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: K-562 cell lysate (input)
Lane 2: HA722972 IP in K-562 cell lysate
Lane 3: Rabbit IgG instead of HA722972 in K-562 cell lysate
Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 41 seconds; ECL: K1801
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