重组Anti-PRMT5抗体[EPR5772] (ab109451)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR5772] to PRMT5
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-PRMT5抗体[EPR5772]
参阅全部 PRMT5 一抗 -
描述
兔单克隆抗体[EPR5772] to PRMT5 -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HEK-293, HepG2, HeLa, and NIH/3T3 cell lysates; mouse and rat brain tissue lysate. ICC/IF: HepG2 and HeLa cells. IHC-P: Human infiltrating duct carcinoma of breast tissue, mouse liver tissue. Flow Cyt (intra): HeLa cells. IP: Mouse brain cell.
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常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
解离常数(KD)
KD = 6.70 x 10 -11 M Learn more about KD -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, PBS, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR5772 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
- Hep G2 nuclear extract lysate (ab14660)
- NIH/3T3 nuclear extract lysate (ab14874)
- Rat brain normal tissue lysate - membrane extract (ab29473)
- Mouse brain tissue lysate - total protein (ab30151)
- Mouse brain tissue lysate - total protein (ab4022)
- Rat brain cerebellum tissue lysate - total protein (ab4032)
- Rat brain cerebrum tissue lysate - total protein (ab4033)
- Mouse brain tissue lysate - total protein (0 days) (ab7188)
- Mouse brain tissue lysate - total protein (14 days) (ab7189)
- Mouse brain tissue lysate - total protein (7 days) (ab7190)
- HEK-293 whole cell lysate (ab7902)
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab109451于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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Flow Cyt (Intra) |
1/100.
For unpurified use at 1/500 - 1/1000.ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (1) |
1/10000 - 1/50000. Detects a band of approximately 72 kDa (predicted molecular weight: 73 kDa).
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IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified use at 1/100 - 1/250. |
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ICC/IF |
1/50.
For purifed use at 1/50For unpurified use at 1/100 - 1/250. |
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IP |
1/30.
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说明 |
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Flow Cyt (Intra)
1/100. For unpurified use at 1/500 - 1/1000.ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/10000 - 1/50000. Detects a band of approximately 72 kDa (predicted molecular weight: 73 kDa). |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/100 - 1/250. |
ICC/IF
1/50. For purifed use at 1/50For unpurified use at 1/100 - 1/250. |
IP
1/30. |
靶标
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功能
Arginine methyltransferase that can both catalyze the formation of omega-N monomethylarginine (MMA) and symmetrical dimethylarginine (sDMA), with a preference for the formation of MMA. Specifically mediates the symmetrical dimethylation of arginine residues in the small nuclear ribonucleoproteins Sm D1 (SNRPD1) and Sm D3 (SNRPD3); such methylation being required for the assembly and biogenesis of snRNP core particles. Methylates SUPT5H. Mono- and dimethylates arginine residues of myelin basic protein (MBP) in vitro. Plays a role in the assembly of snRNP core particles. May play a role in cytokine-activated transduction pathways. Negatively regulates cyclin E1 promoter activity and cellular proliferation. May regulate the SUPT5H transcriptional elongation properties. May be part of a pathway that is connected to a chloride current, possibly through cytoskeletal rearrangement. Methylates histone H2A and H4 'Arg-3' during germ cell development. Methylates histone H3 'Arg-8', which may repress transcription. Methylates the Piwi proteins (PIWIL1, PIWIL2 and PIWIL4), methylation of Piwi proteins being required for the interaction with Tudor domain-containing proteins and subsequent localization to the meiotic nuage. Methylates RPS10. -
组织特异性
Ubiquitous. -
序列相似性
Belongs to the protein arginine N-methyltransferase family. -
翻译后修饰
Disulfide bonds and non-covalent association mediate homooligomers formation. -
细胞定位
Cytoplasm. Nucleus. - Information by UniProt
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数据库链接
- Entrez Gene: 10419 Human
- Entrez Gene: 27374 Mouse
- Entrez Gene: 364382 Rat
- Omim: 604045 Human
- SwissProt: O14744 Human
- SwissProt: Q8CIG8 Mouse
- Unigene: 367854 Human
- Unigene: 196585 Mouse
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别名
- 72 kDa ICln binding protein antibody
- 72 kDa ICln-binding protein antibody
- ANM5_HUMAN antibody
see all
图片
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PRMT5 was immunoprecipitated from 0.35 mg Mouse brain tissue lysate 10 ug with ab109451 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab109451 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: Mouse brain tissue lysate 10 ug
Lane 2: ab109451 IP in Mouse brain tissue lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab109451 in mouse brain tissue lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 100 seconds
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All lanes : Anti-PRMT5 antibody [EPR5772] (ab109451) at 1/50000 dilution (purified)
Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate
Lane 2 : HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 73 kDa
Observed band size: 72 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
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Expression and localization of candidate epigenetic reprogramming factors in human embryos.
Human blastocysts were incubated with primary antibodies for (Panel B) PRMT5 (green), the nuclear DNA stained with DAPI (blue) and visualized by multi-channel confocal microscopy. Note that the expression of PRMT5 primarily localized to the ICM of human blastocysts (indicated by white arrows).
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Anti-PRMT5 antibody [EPR5772] (ab109451) at 1/10000 dilution (purified) + HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 20 µg
Secondary
Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 73 kDa
Observed band size: 72 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
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All lanes : Anti-PRMT5 antibody [EPR5772] (ab109451) at 1/10000 dilution (purified)
Lane 1 : Mouse brain tissue lysate
Lane 2 : Rat brain tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 73 kDa
Observed band size: 72 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
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Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling PRMT5 (green) with purified ab109451 at 1/50.
Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.
Control: Secondary antibody Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/400).
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ICC/IF image of ab109451 (unpurified) stained HepG2 (Human liver hepatocellular carcinoma cell line) cells.
The cells were fixed with 100% methanol (5 min) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab109451, 1/100 dilution) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 µM.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling PRMT5 with purified ab109451 at 1/100.
A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody.
Negative control using PBS instead of primary antibody.
Counterstained with hematoxylin.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human infiltrating duct carcinoma of breast tissue sections labeling PRMT5 with purified ab109451 at 1/100.
A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody.
Negative control using PBS instead of primary antibody.
Counterstained with hematoxylin.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Overlay histogram showing HeLa (Human liver hepatocellular carcinoma cell line) cells stained with ab109451 (unpurified, red line).
The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109451, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monclonal) (1µg/1x106 cells) used under the same conditions.
Acquisition of >5,000 events was performed.
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Equilibrium disassociation constant (KD)
Click here to learn more about KD
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (46)
ab109451 被引用在 46 文献中.
- Li J et al. Oxygen-sensitive methylation of ULK1 is required for hypoxia-induced autophagy. Nat Commun 13:1172 (2022). PubMed: 35246531
- Szewczyk MM et al. PRMT5 regulates ATF4 transcript splicing and oxidative stress response. Redox Biol 51:102282 (2022). PubMed: 35305370
- Dansu DK et al. PRMT5 Interacting Partners and Substrates in Oligodendrocyte Lineage Cells. Front Cell Neurosci 16:820226 (2022). PubMed: 35370564
- Liu H et al. PRMT5 critically mediates TMAO-induced inflammatory response in vascular smooth muscle cells. Cell Death Dis 13:299 (2022). PubMed: 35379776
- Zhang J et al. The PRMT5-LSD1 axis confers Slug dual transcriptional activities and promotes breast cancer progression. J Exp Clin Cancer Res 41:191 (2022). PubMed: 35655230