Name: Anti-PRMT5 antibody [EPR5772]
Brand: RabMAb
SKU: ab109451
Rating: 5 (2 reviews)

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Immunoprecipitation - Anti-PRMT5 antibody [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

Carrier Free PE

製品名

Anti-PRMT5 antibody [EPR5772]
PRMT5 一次抗体製品一覧
製品の詳細

Rabbit monoclonal [EPR5772] to PRMT5
由来種

Rabbit
アプリケーション

適用あり: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details
種交差性

交差種: Mouse, Rat, Human
免疫原

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
ポジティブ･コントロール
- 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.
特記事項
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

製品の状態

Liquid
保存方法

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.
解離定数（K_D 値）

K_D = 6.70 x 10 ^-11 M
Learn more about K_D
バッファー

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, PBS, 0.05% BSA
Concentration information loading...
精製度

Protein A purified
ポリ/モノ

モノクローナル
クローン名

EPR5772
アイソタイプ

IgG
研究分野

The Abpromise guarantee

Abpromise保証は、次のテスト済みアプリケーションにおけるab109451の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション	Abreviews	特記事項
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)	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 1/50 For unpurified use at 1/100 - 1/250.
IP		1/30.

特記事項
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 1/50 For unpurified use at 1/100 - 1/250.
IP 1/30.

機能

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.
組織特異性

Ubiquitous.
配列類似性

Belongs to the protein arginine N-methyltransferase family.
翻訳後修飾

Disulfide bonds and non-covalent association mediate homooligomers formation.
細胞内局在

Cytoplasm. Nucleus.
Target information above from: UniProt accession O14744 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
参照データベース
- 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
別名
- 72 kDa ICln binding protein antibody
- 72 kDa ICln-binding protein antibody
- ANM5_HUMAN antibody
- Histone synthetic lethal 7, S. cerevisiae, homolog of antibody
- Histone-arginine N-methyltransferase PRMT5 antibody
- HMT1 hnRNP methyltransferase like 5 antibody
- HOMOLOG OF; SKB1 antibody
- HRMT1L5 antibody
- IBP72 antibody
- Jak-binding protein 1 antibody
- JBP 1 antibody
- JBP1 antibody
- PRMT 5 antibody
- PRMT5 antibody
- Protein arginine methyltransferase 5 antibody
- Protein arginine N methyltransferase 5 antibody
- Protein arginine N methyltransferase 5 N terminally processed antibody
- Protein arginine N-methyltransferase 5 antibody
- S. POMBE antibody
- S. POMBE HOMOLOG OF; SKB1 antibody
- SHK1 KINASE BINDING PROTEIN 1 antibody
- Shk1 kinase binding protein 1 homolog antibody
- Shk1 kinase-binding protein 1 homolog antibody
- Shk1 kinase/binding protein 1, S. pombe, homolog of antibody
- SKB 1 antibody
- SKB1 antibody
- SKB1 homolog antibody
- SKB1: SKB1 homolog (S. pombe) antibody
- SKB1Hs antibody
see all

Immunoprecipitation - Anti-PRMT5 antibody [EPR5772] (ab109451)

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
Western blot - Anti-PRMT5 antibody [EPR5772] (ab109451)

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.
Immunocytochemistry/ Immunofluorescence - Anti-PRMT5 antibody [EPR5772] (ab109451)Goyal et al PLoS One. 2013 Dec 12;8(12):e82838. doi: 10.1371/journal.pone.0082838. eCollection 2013. Fig 3. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

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).
Western blot - Anti-PRMT5 antibody [EPR5772] (ab109451)

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.
Western blot - Anti-PRMT5 antibody [EPR5772] (ab109451)

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.
Immunocytochemistry/ Immunofluorescence - Anti-PRMT5 antibody [EPR5772] (ab109451)

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).
Immunocytochemistry/ Immunofluorescence - Anti-PRMT5 antibody [EPR5772] (ab109451)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRMT5 antibody [EPR5772] (ab109451)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRMT5 antibody [EPR5772] (ab109451)

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.
Flow Cytometry (Intracellular) - Anti-PRMT5 antibody [EPR5772] (ab109451)

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.
OI-RD Scanning - Anti-PRMT5 antibody [EPR5772] (ab109451)

Equilibrium disassociation constant (K_D)

Click here to learn more about K_D
Anti-PRMT5 antibody [EPR5772] (ab109451)

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

Datasheet download

Download

ab109451 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab109451 は 35 報の論文で使用されています。

Liu X et al. EPZ015666, a selective protein arginine methyltransferase 5 (PRMT5) inhibitor with an antitumour effect in retinoblastoma. Exp Eye Res 202:108286 (2021). PubMed: 33035554
Yang L et al. PRMT5 functionally associates with EZH2 to promote colorectal cancer progression through epigenetically repressing CDKN2B expression. Theranostics 11:3742-3759 (2021). PubMed: 33664859
Mei S et al. PRMT5 promotes progression of endometrioid adenocarcinoma via ERa and cell cycle signaling pathways. J Pathol Clin Res 7:154-164 (2021). PubMed: 33416213
Li WJ et al. Profiling PRMT methylome reveals roles of hnRNPA1 arginine methylation in RNA splicing and cell growth. Nat Commun 12:1946 (2021). PubMed: 33782401
Cai C et al. PRMT5 Enables Robust STAT3 Activation via Arginine Symmetric Dimethylation of SMAD7. Adv Sci (Weinh) 8:2003047 (2021). PubMed: 34026434

View all Publications for this product

レビューと Q&A

Show All レビュー Q&A

レビューを送る質問を送る

1-3 of 3 Abreviews or Q&A

Application

ChIP

Sample

Mouse Cell lysate - whole cell (Mouse NPCs/NSCs)

Specification

Mouse NPCs/NSCs

Detection step

Real-time PCR

Type

Cross-linking (X-ChIP)
Duration of cross-linking step: 60 minute(s) and 0 second(s)
Specification of the cross-linking agent: Bioruptor

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Mr. Nitin Khandelwal

Verified customer

投稿 Jun 19 2015

Application

Western blot

Sample

Human Cell lysate - whole cell (293T, HepG2)

Loading amount

20 µg

Specification

293T, HepG2

Gel Running Conditions

Reduced Denaturing (8%)

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 7% · Temperature: 20°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Mr. Dongil Kim

Verified customer

投稿 Feb 01 2013

Question

samples; HEK293 cells block: 5% milk ab413, ab415: black membrane, protein ladder in white Ab 1/1000 suggestion: use less primary and/or secondary antibody ab109451: lots of bands, Ab 1/1000 suggestion: use less primary antibody (recommended dilution is 1/10,000 to 1/50,000)

Abcam community

Verified customer

Asked on Nov 22 2011

Answer

Thank you for your phone call. I am sorry to hear that these 3 antibodies are not providing satisfactory results. The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. As we discussed over the phone, I would like to offer some suggestions to help optimize the results: For ab413, ab415 I'd suggest using less primary and/or secondary antibody as the membrane shouldn't turn black. This might be an indication of too much antibody on the membrane. For ab109451, most likely using less primary antibody could help with the background as the recommended dilution for the antibody is 1/10,000 to 1/50,000. Should the suggestions not improve the results, please do let me know. In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund. I hope this information is helpful, and I thank you for your cooperation.

Abcam Scientific Support

Answered on Nov 22 2011

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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Key features and details

Related conjugates and formulations

製品の概要

製品名

製品の詳細

由来種

アプリケーション

種交差性

免疫原

ポジティブ･コントロール

特記事項

製品の特性

製品の状態

保存方法

解離定数（KD 値）

バッファー

精製度

ポリ/モノ

クローン名

アイソタイプ

研究分野

関連製品

Alternative Versions

Compatible Secondaries

Isotype control

Positive Controls

Recombinant Protein

アプリケーション

The Abpromise guarantee

ターゲット情報

機能

組織特異性

配列類似性

翻訳後修飾

細胞内局在

参照データベース

別名

画像

プロトコール

データシートおよび資料

Datasheet download

参考文献 (35)

レビューと Q&A

ChIP abreview for Anti-PRMT5 antibody [EPR5772]

Western blot abreview for Anti-PRMT5 antibody [EPR5772]

解離定数（K_D 値）