AB189907

Anti-Pirh2 antibody [EPR18553]

RabMAb
Recombinant
KO Validated
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(4 Publications)

Rabbit Recombinant Monoclonal Pirh2 antibody. Suitable for IP, WB, ICC/IF and reacts with Human, Mouse samples. Cited in 4 publications.

View Alternative Names

ARNIP, CHIMP, PIRH2, RNF199, ZNF363, RCHY1, RING finger and CHY zinc finger domain-containing protein 1, Androgen receptor N-terminal-interacting protein, CH-rich-interacting match with PLAG1, E3 ubiquitin-protein ligase Pirh2, RING finger protein 199, RING-type E3 ubiquitin transferase RCHY1, Zinc finger protein 363, p53-induced RING-H2 protein, hPirh2

8 Images

Immunocytochemistry/ Immunofluorescence - Anti-Pirh2 antibody [EPR18553] (AB189907)

ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Pirh2 antibody [EPR18553] (AB189907)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Pirh2 with ab189907 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing nuclear and cytoplasmic staining on HeLa cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows : -

-ve control 1 : ab189907 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.

-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Pirh2 antibody [EPR18553] (AB189907)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293 (Human epithelial cells from embryonic kidney) cells labeling Pirh2 with ab189907 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing nuclear and cytoplasmic staining on HEK-293 cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows : -

-ve control 1 : ab189907 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.

-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

Supplier Data

Immunoprecipitation - Anti-Pirh2 antibody [EPR18553] (AB189907)

Pirh2 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab189907 at 1/50 dilution.

Western blot was performed from the immunoprecipitate using ab189907 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : HeLa whole cell lysate 10ug (Input).

Lane 2 : ab189907 IP in HeLa whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab189907 in HeLa whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 30 seconds.

All lanes:

Immunoprecipitation - Anti-Pirh2 antibody [EPR18553] (ab189907)

Predicted band size: 30 kDa

Observed band size: 30 kDa

false

Supplier Data

Western blot - Anti-Pirh2 antibody [EPR18553] (AB189907)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Pirh2 antibody [EPR18553] (ab189907) at 1/1000 dilution

Lane 1:

HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate at 10 µg

Lane 2:

HCT 116 (Human colorectal carcinoma cell line) whole cell lysate at 10 µg

Lane 3:

LNCaP (Human prostate cancer cell line) whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 30 kDa

Observed band size: 30 kDa

false

Exposure time: 3min

Lab

Western blot - Anti-Pirh2 antibody [EPR18553] (AB189907)

Lanes 1-4 : Merged signal (red and green). Green - ab189907 observed at 30 kDa. Red - loading control ab8245 observed at 37 kDa.

ab189907 Anti-Pirh2 antibody [EPR18553] was shown to specifically react with RCHY1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265478 (knockout cell lysate ab258171) was used. Wild-type and RCHY1 knockout samples were subjected to SDS-PAGE. ab189907 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Pirh2 antibody [EPR18553] (ab189907) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

RCHY1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human RCHY1 (Pirh2) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-rchy1-pirh2-knockout-hela-cell-line-ab265478'>ab265478</a>)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Predicted band size: 30 kDa

Observed band size: 30 kDa

false

Supplier Data

Western blot - Anti-Pirh2 antibody [EPR18553] (AB189907)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Pirh2 antibody [EPR18553] (ab189907) at 1/1000 dilution

Lanes 1 and 3:

Human fetal heart lysate at 10 µg

Lane 2:

Human fetal kidney lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 30 kDa

Observed band size: 30 kDa

false

Exposure time: 30s

Supplier Data

Western blot - Anti-Pirh2 antibody [EPR18553] (AB189907)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Pirh2 antibody [EPR18553] (ab189907) at 1/1000 dilution

Lane 1:

Mouse kidney lysate at 10 µg

Lane 2:

Mouse spleen lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 30 kDa

Observed band size: 30 kDa

false

Exposure time: 3min

Supplier Data

Western blot - Anti-Pirh2 antibody [EPR18553] (AB189907)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Pirh2 antibody [EPR18553] (ab189907) at 1/1000 dilution

Lane 1:

RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 2:

NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 30 kDa

Observed band size: 30 kDa

false

Exposure time: 3min

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR18553

Isotype

IgG

Carrier free

Reacts with

Mouse, Human

Applications

IP, WB, ICC/IF

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "1/50", "IP-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/500", "ICCIF-species-notes": "" }, "Mouse": { "IP-species-checked": "guaranteed", "IP-species-dilution-info": "", "IP-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "ICCIF-species-checked": "guaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "" } } }

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Product details

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Pirh2 also known as RCHY1 is a protein with E3 ubiquitin ligase activity. It has a molecular mass of approximately 34 kDa. Pirh2 targets proteins for degradation by the ubiquitin-proteasome system regulating the levels of proteins like p53. Expression of Pirh2 occurs in various tissues including liver and heart. It helps maintain cellular protein homeostasis by marking specific substrates for degradation.

Biological function summary

Pirh2 influences the regulation of protein stability by tagging proteins for degradation. Besides p53 it acts on other substrates and engages in various cellular processes. Pirh2 operates independently and its presence affects cellular responses to stress and DNA damage. It modulates cell cycle progression and apoptosis impacting cell fate decisions.

Pathways

Pirh2 plays an important role in the p53 signaling pathway an essential cell regulatory network. By mediating the ubiquitination of p53 Pirh2 affects cell survival and division under stress conditions. Interaction with proteins such as Mdm2 further regulates p53 activity impacting the cell's ability to respond to DNA damage and carcinogen exposure.

Research connects Pirh2 with the development of cancers. Its interaction with the tumor suppressor protein p53 poses effects on tumorigenesis by altering p53 stability and function. Overexpression of Pirh2 has connections to several cancer types including breast and liver cancers. These associations suggest Pirh2 as a potential target for therapeutic interventions that aim at modulating the p53 pathway to treat cancer.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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Target data

E3 ubiquitin-protein ligase that mediates ubiquitination of target proteins, including p53/TP53, TP73, HDAC1 and CDKN1B (PubMed : 16914734, PubMed : 17721809, PubMed : 18006823, PubMed : 19043414, PubMed : 19483087, PubMed : 21994467). Mediates ubiquitination and degradation of p53/TP53; preferentially acts on tetrameric p53/TP53 (PubMed : 19043414, PubMed : 19483087). Catalyzes monoubiquitinates the translesion DNA polymerase POLH (PubMed : 21791603). Involved in the ribosome-associated quality control (RQC) pathway, which mediates the extraction of incompletely synthesized nascent chains from stalled ribosomes : RCHY1 acts downstream of NEMF and recognizes CAT tails associated with stalled nascent chains, leading to their ubiquitination and degradation (PubMed : 33909987).. Isoform 4. Has no E3 ubiquitin-protein ligase activity.

See full target information RCHY1

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Publications (4)

Recent publications for all applications. Explore the full list and refine your search

Cell reports. Medicine 4:101277 PubMed37944531

2023

Personalized drug screening in patient-derived organoids of biliary tract cancer and its clinical application.

Applications

Unspecified application

Species

Unspecified reactive species

Xiaoxue Ren,Mingle Huang,Weixiang Weng,Yubin Xie,Yifan Wu,Shenghua Zhu,Ying Zhang,Dongming Li,Jiaming Lai,Shunli Shen,Jie Lin,Ming Kuang,Xiaoxing Li,Jun Yu,Lixia Xu

Cell research 33:835-850 PubMed37726403

2023

Low glucose metabolite 3-phosphoglycerate switches PHGDH from serine synthesis to p53 activation to control cell fate.

Applications

Unspecified application

Species

Unspecified reactive species

Yu-Qing Wu,Chen-Song Zhang,Jinye Xiong,Dong-Qi Cai,Chen-Zhe Wang,Yu Wang,Yan-Hui Liu,Yu Wang,Yiming Li,Jian Wu,Jianfeng Wu,Bin Lan,Xuefeng Wang,Siwei Chen,Xianglei Cao,Xiaoyan Wei,Hui-Hui Hu,Huiling Guo,Yaxin Yu,Abdul Ghafoor,Changchuan Xie,Yaying Wu,Zheni Xu,Cixiong Zhang,Mingxia Zhu,Xi Huang,Xiufeng Sun,Shu-Yong Lin,Hai-Long Piao,Jianyin Zhou,Sheng-Cai Lin

Cancer management and research 12:13149-13159 PubMed33376406

2020

Pyrrolidine Dithiocarbamate Facilitates Arsenic Trioxide Against Pancreatic Cancer via Perturbing Ubiquitin-Proteasome Pathway.

Applications

Unspecified application

Species

Unspecified reactive species

Simin Yu,Ning Wu,Jianmin Zhu,Ying Liu,Jinbin Han

Journal of experimental & clinical cancer research 39:190 PubMed32938489

2020

IMP3 accelerates the progression of prostate cancer through inhibiting PTEN expression in a SMURF1-dependent way.

Applications

Unspecified application

Species

Unspecified reactive species

Xiang Zhang,Dawei Wang,Boke Liu,Xingwei Jin,Xianjin Wang,Junwei Pan,Weichao Tu,Yuan Shao

View all publications

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Anti-Pirh2 antibody [EPR18553]

Immunocytochemistry/ Immunofluorescence - Anti-Pirh2 antibody [EPR18553] (AB189907)

Immunocytochemistry/ Immunofluorescence - Anti-Pirh2 antibody [EPR18553] (AB189907)

Immunoprecipitation - Anti-Pirh2 antibody [EPR18553] (AB189907)

Western blot - Anti-Pirh2 antibody [EPR18553] (AB189907)

Western blot - Anti-Pirh2 antibody [EPR18553] (AB189907)

Western blot - Anti-Pirh2 antibody [EPR18553] (AB189907)

Western blot - Anti-Pirh2 antibody [EPR18553] (AB189907)

Western blot - Anti-Pirh2 antibody [EPR18553] (AB189907)

Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Alternative Products

Primary Antibodies

Complementary Products

Primary Antibodies

Primary Antibodies

Primary Antibodies

Primary Antibodies

Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage duration

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Aliquoting information

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Publications (4)

Product promise