Invitrogen

PRMT5 Monoclonal Antibody (PRMT5-21)

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This Antibody was verified by KnockDown to ensure that the antibody binds to the antigen stated. View Details

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PRMT5 Antibody in Western Blot (WB) — Knockdown of PRMT5 was achieved by transfecting HeLa cells with PRMT5 specific siRNAs (Silencer® select Product # s20377). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1% SDS) from the PRMT5 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with PRMT5 Monoclonal Antibody (PRMT1) (Product # MA1-25470, 2 µg/mL) and Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to PRMT5.

PRMT5 Antibody in Western Blot (WB) — Western blot analysis was performed on modified whole cell extracts (30 µg lysate) of HeLa (Lane 1), T-47D (Lane 2), Caco-2 (Lane 3), HepG2 (Lane 4), PC-3 (Lane 5) and NIH/3T3 (Lane 6). The blot was probed with Anti-PRMT5 Monoclonal Antibody (PRMT5-21) (Product # MA1-25470, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). A 72 kDa band corresponding to PRMT5 was observed in all human and mouse cell line(s) tested.

PRMT5 Antibody in ChIP assay (ChIP) — Enrichment of endogenous PRMT5 protein at specific gene loci using Anti-PRMT5 Antibody: Chromatin Immunoprecipitation (ChIP) was performed using Anti-PRMT5 Mouse Monoclonal antibody (Product # MA1-25470, 5 µg) on sheared chromatin from 2 million HCT 116 cells using the MAGnify ChIP System (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR with PCR primer pairs over the promoters of ADIPOQ, BIRC3, CCNE1, FOXP1 (positive) and SAT2 satellite repeats (negative). Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

PRMT5 Antibody in Relative expression — Antibody specificity was demonstrated by detection of enrichment of the target protein at specific gene loci. Chromatin Immunoprecipitation (ChIP) was performed using Anti-PRMT5 monoclonal antibody (Product # MA1-25470) using PCR primer pairs over the promoters of ADIPOQ, BIRC3, CCNE1, FOXP1 (positive) and SAT2 satellite repeats (negative). {RE}

PRMT5 Antibody in KnockDown — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. HeLa cells were transfected with PRMT5 siRNA and reduction of signal was observed in Western Blot using PRMT5 Monoclonal Antibody (PRMT521) (Product # MA1-25470). {KD}

Product Details

Tested Applications

Dilution

ChIP assay (ChIP)

2.5 µg/10^6 cells

ELISA (ELISA)

Assay Dependent

Western Blot (WB)

2 µg/mL

Product Specifications

Species Reactivity

Bovine, Dog, Hamster, Human, Mouse, Non-human primate, Rat

Host / Isotype

Mouse / IgG2a

Class

Monoclonal

Type

Antibody

Clone

PRMT5-21

Immunogen

Fusion of mouse myeloma cells and splenocytes from mice immunized with PRMT5 containing the C-terminal region of the Human PRMT5 (residues 315-637).

Conjugate

Unconjugated

Form

Liquid

Concentration

2 mg/mL

Purification

Protein A

Storage buffer

0.01M PBS, pH 7.4

Contains

15mM sodium azide

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RRID

AB_2253071

Product Specific Information

MA1-25470 detects PRMT5 from human, mouse, rat, bovine, canine, hamster, and monkey samples.

MA1-25470 has been successfully used in Western blot and ELISA procedures.

The MA1-25470 immunogen is a fusion of mouse myeloma cells and splenocytes from mice immunized with PRMT5 containing the C-terminal region of the Human PRMT5 (residues 315-637).

Store at 4°C short term. For extended storage, aliquot and store at -20°C, avoiding freeze/thaw cycles. Working dilutions should be discarded if not used within 12 hours.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: 72 kDa ICln-binding protein; Histone-arginine N-methyltransferase PRMT5; HMT1 hnRNP methyltransferase-like 5; HRMT1L5; IBP72; Jak-binding protein 1; JBP1; protein arginine methyltransferase 5; Protein arginine N-methyltransferase 5; Protein arginine N-methyltransferase 5, N-terminally processed; protein arginine N-methyltransferase 5-like; Shk1 kinase-binding protein 1 homolog; SKB1; SKB1 homolog; SKB1Hs

Gene Aliases: BOS_10182; HRMT1L5; I79_013398; IBP72; JBP1; PRMT5; SKB1; SKB1Hs

UniProt ID: (Human) O14744, (Bovine) A7YW45, (Mouse) Q8CIG8

Entrez Gene ID: (Human) 10419, (Bovine) 515594, (Dog) 480242, (Mouse) 27374, (Rat) 364382, (Chinese hamster) 100768836

Molecular Function: signaling molecule

spliceosomal snRNP assembly DNA-templated transcription, termination regulation of transcription, DNA-templated regulation of mitotic nuclear division cell proliferation peptidyl-arginine methylation peptidyl-arginine methylation, to symmetrical-dimethyl arginine circadian regulation of gene expression peptidyl-arginine N-methylation endothelial cell activation histone H4-R3 methylation regulation of DNA methylation negative regulation of cell differentiation positive regulation of oligodendrocyte differentiation regulation of ERK1 and ERK2 cascade Golgi ribbon formation liver regeneration regulation of signal transduction by p53 class mediator negative regulation of nucleic acid-templated transcription positive regulation of adenylate cyclase-inhibiting dopamine receptor signaling pathway transcription, DNA-templated negative regulation of transcription from RNA polymerase II promoter protein methylation chromatin modification methylation histone arginine methylation rhythmic process

Process(es)

core promoter sequence-specific DNA binding transcription corepressor activity protein binding methyltransferase activity methyl-CpG binding histone-arginine N-methyltransferase activity protein-arginine N-methyltransferase activity protein complex binding protein-arginine omega-N symmetric methyltransferase activity ribonucleoprotein complex binding histone methyltransferase activity (H4-R3 specific) protein heterodimerization activity rRNA (adenine-N6,N6-)-dimethyltransferase activity chromatin binding protein C-terminal carboxyl O-methyltransferase activity tRNA (guanine-N2-)-methyltransferase activity C-methyltransferase activity N-methyltransferase activity O-methyltransferase activity S-methyltransferase activity RNA methyltransferase activity mRNA methyltransferase activity tRNA methyltransferase activity protein methyltransferase activity 2-polyprenyl-6-methoxy-1,4-benzoquinone methyltransferase activity rRNA methyltransferase activity rRNA (uridine-2'-O-)-methyltransferase activity rRNA (adenine-N6-)-methyltransferase activity DNA-methyltransferase activity tRNA (guanine-N1-)-methyltransferase activity rRNA (cytosine-C5-)-methyltransferase activity selenocysteine methyltransferase activity arginine N-methyltransferase activity lysine N-methyltransferase activity protein-lysine N-methyltransferase activity tRNA (uracil) methyltransferase activity tRNA (guanine) methyltransferase activity tRNA (adenine) methyltransferase activity tRNA (cytosine) methyltransferase activity tRNA (cytosine-5-)-methyltransferase activity tRNA (adenine-N1-)-methyltransferase activity rRNA (adenine) methyltransferase activity rRNA (cytosine) methyltransferase activity rRNA (guanine) methyltransferase activity rRNA (uridine) methyltransferase activity transferase activity protein C-terminal leucine carboxyl O-methyltransferase activity 1-phenanthrol methyltransferase activity protein-arginine N5-methyltransferase activity methylarsonite methyltransferase activity dimethylarsinite methyltransferase activity 4,5-dihydroxybenzo(a)pyrene methyltransferase activity 1-hydroxypyrene methyltransferase activity 1-hydroxy-6-methoxypyrene methyltransferase activity demethylmenaquinone methyltransferase activity cobalt-precorrin-6B C5-methyltransferase activity cobalt-precorrin-7 C15-methyltransferase activity cobalt-precorrin-5B C1-methyltransferase activity cobalt-precorrin-3 C17-methyltransferase activity dimethylamine methyltransferase activity hydroxyneurosporene-O-methyltransferase activity tRNA (adenine-57, 58-N(1)-) methyltransferase activity methylamine-specific methylcobalamin:coenzyme M methyltransferase activity trimethylamine methyltransferase activity methanol-specific methylcobalamin:coenzyme M methyltransferase activity monomethylamine methyltransferase activity P-methyltransferase activity Se-methyltransferase activity 2-phytyl-1,4-naphthoquinone methyltransferase activity tRNA (uracil-2'-O-)-methyltransferase activity tRNA (cytosine-2'-O-)-methyltransferase activity phosphomethylethanolamine N-methyltransferase activity tRNA (cytosine-3-)-methyltransferase activity rRNA (cytosine-2'-O-)-methyltransferase activity rRNA (cytosine-N4-)-methyltransferase activity trihydroxyferuloyl spermidine O-methyltransferase activity