PRMT7 Antibody [F2F1] | Primary Antibodies

Application: Reactivity: Human

Lane 1: HepG2, Lane 2: Hela, Lane 3: 293, Lane 4: MCF7

Usage Information

Dilution
WB1:1000 - 1:10000 IP1:100 - 1:250

Application
WB, IP

Reactivity
Human

Source
Rabbit Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW
78 kDa

Positive Control	HepG2 cells; HeLa cells; 293 cells; MCF7 cells
Negative Control

Experimental Methods

WB
Experimental Protocol: Sample preparation 1. Tissue: Lyse the tissue sample by adding an appropriate volume of ice-cold RIPA/NP-40 Lysis Buffer (containing Protease Inhibitor Cocktail)，and homogenize the tissue at a low temperature or lyse it by sonication on ice, then incubate on ice for 30 minutes. 2. Adherent cell: Aspirate the culture medium and wash the cells with ice-cold PBS twice. Lyse the cells by adding an appropriate volume of RIPA/NP-40 Lysis Buffer (containing Protease Inhibitor Cocktail) , sonicate to lyse the cells, and incubate on ice for 30 minutes. 3. Suspension cell: Transfer the culture medium to a pre-cooled centrifuge tube. Centrifuge and aspirate the supernatant. Wash the cells with ice-cold PBS twice. Lyse the cells by adding an appropriate volume of RIPA/NP-40 Lysis Buffer (containing Protease Inhibitor Cocktail) , sonicate to lyse the cells, and incubate on ice for 30 minutes. 4. Place the lysate into a pre-cooled microcentrifuge tube. Centrifuge at 4°C for 15 min. Collect the supernatant; 5. Remove a small volume of lysate to determine the protein concentration; 6. Combine the lysate with protein loading buffer. Boil 20 µL sample under 95-100°C for 5 min. Centrifuge for 5 min after cool down on ice. Electrophoretic separation 1. According to the concentration of extracted protein, load appropriate amount of protein sample and marker onto SDS-PAGE gels for electrophoresis. Recommended separating gel (lower gel) concentration: 10%. Reference Table for Selecting SDS-PAGE Separation Gel Concentrations 2. Power up 80V for 30 minutes. Then the power supply is adjusted (110 V~150 V), the Marker is observed, and the electrophoresis can be stopped when the indicator band of the predyed protein Marker where the protein is located is properly separated. (Note that the current should not be too large when electrophoresis, too large current (more than 150 mA) will cause the temperature to rise, affecting the result of running glue. If high currents cannot be avoided, an ice bath can be used to cool the bath.) Transfer membrane 1. Take out the converter, soak the clip and consumables in the pre-cooled converter; 2. Activate PVDF membrane with methanol for 1 min and rinse with transfer buffer; 3. Install it in the order of "black edge of clip - sponge - filter paper - filter paper - glue -PVDF membrane - filter paper - filter paper - sponge - white edge of clip"; 4. The protein was electrotransferred to PVDF membrane. ( 0.45 µm PVDF membrane is recommended ) Reference Table for Selecting PVDF Membrane Pore Size Specifications Recommended conditions for wet transfer: 200 mA, 120 min. ( Note that the transfer conditions can be adjusted according to the protein size. For high-molecular-weight proteins, a higher current and longer transfer time are recommended. However, ensure that the transfer tank remains at a low temperature to prevent gel melting.) Block 1. After electrotransfer, wash the film with TBST at room temperature for 5 minutes; 2. Incubate the film in the blocking solution for 1 hour at room temperature; 3. Wash the film with TBST for 3 times, 5 minutes each time. Antibody incubation 1. Use primary antibody dilution buffer to prepare the primary antibody working liquid (recommended dilution ratio for primary antibody 1:2000), gently shake and incubate with the film at 4°C overnight; 2. Wash the film with TBST 3 times, 5 minutes each time; 3. Add the secondary antibody to the blocking solution and incubate with the film gently at room temperature for 1 hour; 4. After incubation, wash the film with TBST 3 times for 5 minutes each time. Antibody staining 1. Add the prepared ECL luminescent substrate (or select other color developing substrate according to the second antibody) and mix evenly; 2. Incubate with the film for 1 minute, remove excess substrate (keep the film moist), wrap with plastic film, and expose in the imaging system.

WB

Experimental Protocol:

Sample preparation

1. Tissue: Lyse the tissue sample by adding an appropriate volume of ice-cold RIPA/NP-40 Lysis Buffer (containing Protease Inhibitor Cocktail)，and homogenize the tissue at a low temperature or lyse it by sonication on ice, then incubate on ice for 30 minutes.

2. Adherent cell: Aspirate the culture medium and wash the cells with ice-cold PBS twice. Lyse the cells by adding an appropriate volume of RIPA/NP-40 Lysis Buffer (containing Protease Inhibitor Cocktail) , sonicate to lyse the cells, and incubate on ice for 30 minutes.

3. Suspension cell: Transfer the culture medium to a pre-cooled centrifuge tube. Centrifuge and aspirate the supernatant. Wash the cells with ice-cold PBS twice. Lyse the cells by adding an appropriate volume of RIPA/NP-40 Lysis Buffer (containing Protease Inhibitor Cocktail) , sonicate to lyse the cells, and incubate on ice for 30 minutes.

4. Place the lysate into a pre-cooled microcentrifuge tube. Centrifuge at 4°C for 15 min. Collect the supernatant;

5. Remove a small volume of lysate to determine the protein concentration;

6. Combine the lysate with protein loading buffer. Boil 20 µL sample under 95-100°C for 5 min. Centrifuge for 5 min after cool down on ice.

Electrophoretic separation

1. According to the concentration of extracted protein, load appropriate amount of protein sample and marker onto SDS-PAGE gels for electrophoresis. Recommended separating gel (lower gel) concentration: 10%. Reference Table for Selecting SDS-PAGE Separation Gel Concentrations

2. Power up 80V for 30 minutes. Then the power supply is adjusted (110 V~150 V), the Marker is observed, and the electrophoresis can be stopped when the indicator band of the predyed protein Marker where the protein is located is properly separated. (Note that the current should not be too large when electrophoresis, too large current (more than 150 mA) will cause the temperature to rise, affecting the result of running glue. If high currents cannot be avoided, an ice bath can be used to cool the bath.)

Transfer membrane

1. Take out the converter, soak the clip and consumables in the pre-cooled converter;

2. Activate PVDF membrane with methanol for 1 min and rinse with transfer buffer;

3. Install it in the order of "black edge of clip - sponge - filter paper - filter paper - glue -PVDF membrane - filter paper - filter paper - sponge - white edge of clip";

4. The protein was electrotransferred to PVDF membrane. ( 0.45 µm PVDF membrane is recommended ) Reference Table for Selecting PVDF Membrane Pore Size Specifications

Recommended conditions for wet transfer: 200 mA, 120 min.

( Note that the transfer conditions can be adjusted according to the protein size. For high-molecular-weight proteins, a higher current and longer transfer time are recommended. However, ensure that the transfer tank remains at a low temperature to prevent gel melting.)

Block

1. After electrotransfer, wash the film with TBST at room temperature for 5 minutes;

2. Incubate the film in the blocking solution for 1 hour at room temperature;

3. Wash the film with TBST for 3 times, 5 minutes each time.

Antibody incubation

1. Use primary antibody dilution buffer to prepare the primary antibody working liquid (recommended dilution ratio for primary antibody 1:2000), gently shake and incubate with the film at 4°C overnight;

2. Wash the film with TBST 3 times, 5 minutes each time;

3. Add the secondary antibody to the blocking solution and incubate with the film gently at room temperature for 1 hour;

4. After incubation, wash the film with TBST 3 times for 5 minutes each time.

Antibody staining

1. Add the prepared ECL luminescent substrate (or select other color developing substrate according to the second antibody) and mix evenly;

2. Incubate with the film for 1 minute, remove excess substrate (keep the film moist), wrap with plastic film, and expose in the imaging system.

Datasheet & SDS

Biological Description

Specificity
PRMT7 Antibody (Rabbit mAb) [F2F1] detects endogenous levels of total PRMT7 protein.

Subcellular Location
Cytoplasm, Nucleus

Uniprot ID
Q9NVM4

Clone
F2F1

Synonym(s)
KIAA1933, PRMT7, Protein arginine N-methyltransferase 7, Histone-arginine N-methyltransferase PRMT7, [Myelin basic protein]-arginine N-methyltransferase PRMT7

Background
PRMT7 is a type III protein arginine methyltransferase that catalyzes predominantly monomethylation of arginine residues on histone and non‑histone substrates and acts as a pleiotropic regulator of gene expression, stemness, stress responses, and neuromuscular development through both direct chromatin modification and crosstalk with other PRMTs, especially PRMT5. The enzyme is built around a duplicated PRMT core with two AdoMet-binding domains arranged in tandem, but only one catalytic site is active, conferring unique substrate specificity and kinetic properties within the PRMT family and favoring MMA formation that can be further converted to symmetric dimethylarginine by PRMT5. PRMT7 methylates arginine residues in glycine–arginine‑rich stretches of histones and snRNP proteins and contributes to the generation of H4R3me2s and Sm protein SDMA indirectly by activating PRMT5 or creating a priming MMA state, which influences chromatin accessibility, snRNP core assembly, and RNA processing capacity. At chromatin, PRMT7‑dependent H4R3 symmetric dimethylation has been linked to repression of DNA damage repair genes such as APEX2, POLD1, and POLD2 and to antagonism of MLL4‑mediated H3K4 methylation at neuron‑specific promoters, positioning PRMT7 as a negative regulator of subsets of repair pathways and neuronal differentiation programs. PRMT7 is enriched in embryonic stem cells, germ cells, and multiple cancer stem cell populations, where its activity supports self‑renewal and stemness by repressing differentiation‑promoting loci, modulating miRNA networks such as the miR‑24‑2 cluster, and maintaining proper levels of SDMA‑marked chromatin and RNA‑binding proteins. In cancer models, altered PRMT7 expression influences proliferation, migration, and stress tolerance: PRMT7 overexpression promotes tumor growth and survival under genotoxic or metabolic stress, while PRMT7 knockdown reduces clonogenicity, induces G1 arrest and differentiation, and sensitizes cells to DNA damaging agents, leading to increasing interest in PRMT7 as a drug target and the recent identification of dual PRMT7/9 inhibitors with defined binding modes.

Background

PRMT7 is a type III protein arginine methyltransferase that catalyzes predominantly monomethylation of arginine residues on histone and non‑histone substrates and acts as a pleiotropic regulator of gene expression, stemness, stress responses, and neuromuscular development through both direct chromatin modification and crosstalk with other PRMTs, especially PRMT5. The enzyme is built around a duplicated PRMT core with two AdoMet-binding domains arranged in tandem, but only one catalytic site is active, conferring unique substrate specificity and kinetic properties within the PRMT family and favoring MMA formation that can be further converted to symmetric dimethylarginine by PRMT5. PRMT7 methylates arginine residues in glycine–arginine‑rich stretches of histones and snRNP proteins and contributes to the generation of H4R3me2s and Sm protein SDMA indirectly by activating PRMT5 or creating a priming MMA state, which influences chromatin accessibility, snRNP core assembly, and RNA processing capacity. At chromatin, PRMT7‑dependent H4R3 symmetric dimethylation has been linked to repression of DNA damage repair genes such as APEX2, POLD1, and POLD2 and to antagonism of MLL4‑mediated H3K4 methylation at neuron‑specific promoters, positioning PRMT7 as a negative regulator of subsets of repair pathways and neuronal differentiation programs. PRMT7 is enriched in embryonic stem cells, germ cells, and multiple cancer stem cell populations, where its activity supports self‑renewal and stemness by repressing differentiation‑promoting loci, modulating miRNA networks such as the miR‑24‑2 cluster, and maintaining proper levels of SDMA‑marked chromatin and RNA‑binding proteins. In cancer models, altered PRMT7 expression influences proliferation, migration, and stress tolerance: PRMT7 overexpression promotes tumor growth and survival under genotoxic or metabolic stress, while PRMT7 knockdown reduces clonogenicity, induces G1 arrest and differentiation, and sensitizes cells to DNA damaging agents, leading to increasing interest in PRMT7 as a drug target and the recent identification of dual PRMT7/9 inhibitors with defined binding modes.

References
https://pubmed.ncbi.nlm.nih.gov/34440512/ https://pubmed.ncbi.nlm.nih.gov/34712331/

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%

PRMT7 Antibody (Rabbit mAb) [F2F1]

Usage Information

Experimental Methods

Datasheet & SDS

Biological Description

Tech Support