Recombinant Anti-Pirh2 antibody [EPR18553] KO Tested (ab189907)

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Western blot - Anti-Pirh2 antibody [EPR18553] (ab189907)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18553] to Pirh2
Suitable for: ICC/IF, IP, WB
Knockout validated
Reacts with: Mouse, Human

Carrier Free

Product name

Anti-Pirh2 antibody [EPR18553]
See all Pirh2 primary antibodies
Description

Rabbit monoclonal [EPR18553] to Pirh2
Host species

Rabbit
Tested applications

Suitable for: ICC/IF, IP, WBmore details
Species reactivity

Reacts with: Mouse, Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HeLa, HepG2, Daudi, HEK-293, HCT 116, LNCaP, RAW 264.7 and NIH/3T3 whole cell lysates; Human fetal heart, fetal kidney and fetal spleen lysates; Mouse kidney and spleen lysates. ICC/IF: HeLa and HEK-293 cells. IP: HeLa whole cell lysate.
General notes
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.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18553
Isotype

IgG
Research areas

Alternative Versions
- Anti-Pirh2 antibody [EPR18553] - BSA and Azide free (ab251001)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
- Human RCHY1 (Pirh2) knockout HeLa cell line (ab265478)
KO cell lysates
- Human RCHY1 (Pirh2) knockout HeLa cell lysate (ab258171)
Positive Controls
- VeriBlot for IP Detection Reagent (HRP) (ab131366)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab189907 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
ICC/IF		1/500.
IP		1/50.
WB		1/1000. Detects a band of approximately 30 kDa (predicted molecular weight: 30 kDa).

Notes
ICC/IF 1/500.
IP 1/50.
WB 1/1000. Detects a band of approximately 30 kDa (predicted molecular weight: 30 kDa).

Function

Mediates E3-dependent ubiquitination and proteasomal degradation of target proteins, including p53/TP53, HDAC1 and CDKN1B. Preferentially acts on tetrameric p53/TP53. Contributes to the regulation of CDKN1B and p53/TP53 levels, and thereby contributes to the regulation of the cell cycle progression. Increases AR transcription factor activity.
Pathway

Protein modification; protein ubiquitination.
Sequence similarities

Contains 1 CHY-type zinc finger.
Contains 1 CTCHY-type zinc finger.
Contains 1 RING-type zinc finger.
Post-translational
modifications

Subject to ubiquitination and proteasomal degradation. Interaction with PLAGL2 or KAT5 enhances protein stability.
Cellular localization

Nucleus. Nucleus speckle. Cytoplasm.
Target information above from: UniProt accession Q96PM5 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 25898 Human
- Entrez Gene: 68098 Mouse
- Omim: 607680 Human
- SwissProt: Q96PM5 Human
- SwissProt: Q9CR50 Mouse
- Unigene: 48297 Human
- Unigene: 159453 Mouse
Alternative names
- Androgen receptor N terminal interacting protein antibody
- Androgen receptor N-terminal-interacting protein antibody
- ARNIP antibody
- CH-rich-interacting match with PLAG1 antibody
- CHIMP antibody
- E3 ubiquitin-protein ligase Pirh2 antibody
- hARNIP antibody
- hPirh2 antibody
- p53 induced protein with a RING H2 domain antibody
- p53-induced RING-H2 protein antibody
- PIRH2E antibody
- PIRH2F antibody
- PRO1996 antibody
- RCHY1 antibody
- Ring finger and CHY zinc finger domain containing 1 E3 ubiquitin protein ligase antibody
- RING finger and CHY zinc finger domain-containing protein 1 antibody
- RING finger protein 199 antibody
- RNF199 antibody
- ZCHY antibody
- ZFP 363 antibody
- zinc finger CHY type antibody
- Zinc finger protein 363 antibody
- ZN363_HUMAN antibody
- ZNF363 antibody
see all

Western blot - Anti-Pirh2 antibody [EPR18553] (ab189907)

All lanes : Anti-Pirh2 antibody [EPR18553] (ab189907) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : RCHY1 knockout HeLa cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : Daudi cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 30 kDa

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.
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.
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.
Western blot - Anti-Pirh2 antibody [EPR18553] (ab189907)

All lanes : Anti-Pirh2 antibody [EPR18553] (ab189907) at 1/1000 dilution

Lane 1 : HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate
Lane 2 : HCT 116 (Human colorectal carcinoma cell line) whole cell lysate
Lane 3 : LNCaP (Human prostate cancer cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 30 kDa
Observed band size: 30 kDa

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-Pirh2 antibody [EPR18553] (ab189907)

All lanes : Anti-Pirh2 antibody [EPR18553] (ab189907) at 1/1000 dilution

Lanes 1 & 3 : Human fetal heart lysate
Lane 2 : Human fetal kidney lysate

Lysates/proteins at 10 µg per lane.

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

Exposure time: 30 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-Pirh2 antibody [EPR18553] (ab189907)

All lanes : Anti-Pirh2 antibody [EPR18553] (ab189907) at 1/1000 dilution

Lane 1 : Mouse kidney lysate
Lane 2 : Mouse spleen lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 30 kDa
Observed band size: 30 kDa

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-Pirh2 antibody [EPR18553] (ab189907)

All lanes : 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
Lane 2 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 30 kDa
Observed band size: 30 kDa

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
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.
Anti-Pirh2 antibody [EPR18553] (ab189907)

Western blot protocols
Immunoprecipitation protocols
Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

SDS download

Country/Region

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Datasheet download

Download

Publishing research using ab189907? Please let us know so that we can cite the reference in this datasheet.

ab189907 has been referenced in 2 publications.

Zhang X et al. IMP3 accelerates the progression of prostate cancer through inhibiting PTEN expression in a SMURF1-dependent way. J Exp Clin Cancer Res 39:190 (2020). PubMed: 32938489
Yu S et al. Pyrrolidine Dithiocarbamate Facilitates Arsenic Trioxide Against Pancreatic Cancer via Perturbing Ubiquitin-Proteasome Pathway. Cancer Manag Res 12:13149-13159 (2020). PubMed: 33376406

Customer reviews and Q&As

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

KO cell lines

KO cell lysates

Positive Controls

Applications

The Abpromise guarantee

Target

Function

Pathway

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (2)

Customer reviews and Q&As

Post-translational
modifications