JavaScript is disabled in your browser. Please enable JavaScript to view this website.
AB258171

Human RCHY1 (Pirh2) knockout HeLa cell lysate

Be the first to review this product! Submit a review

|

(0 Publication)

RCHY1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 11 bp deletion in exon1 and 1 bp insertion in exon1 and 4 bp deletion in exon1.
5 Images
Western blot - Human RCHY1 (Pirh2) knockout HeLa cell lysate (AB258171)
  • WB

Lab

Western blot - Human RCHY1 (Pirh2) knockout HeLa cell lysate (AB258171)

Lane 1 : Wild-type HeLa cell lysate (20 μg)

Lane 2 : RCHY1 knockout HeLa cell lysate (20 μg)

Lane 3 : HepG2 cell lysate (20 μg)

Lane 4 : Daudi cell lysate (20 μg)

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] (<a href='/en-us/products/primary-antibodies/pirh2-antibody-epr18553-ab189907'>ab189907</a>) 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

Western blot - Human RCHY1 (Pirh2) knockout HeLa cell lysate (AB258171)
  • WB

Lab

Western blot - Human RCHY1 (Pirh2) knockout HeLa cell lysate (AB258171)

Lane 1 : Wild-type HeLa cell lysate (20 μg)

Lane 2 : RCHY1 knockout HeLa cell lysate (20 μg)

Lane 3 : HepG2 cell lysate (20 μg)

Lane 4 : Daudi cell lysate (20 μg)

Lanes 1-4 : Merged signal (red and green). Green - ab189247 observed at 30 kDa. Red - loading control ab8245 observed at 37 kDa.

ab189247 Recombinant Anti-Pirh2 antibody [EPR14980] 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. ab189247 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 [EPR14980] (<a href='/en-us/products/primary-antibodies/pirh2-antibody-epr14980-ab189247'>ab189247</a>) 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

Sanger Sequencing - Human RCHY1 (Pirh2) knockout HeLa cell lysate (AB258171)
  • Sanger seq

Unknown

Sanger Sequencing - Human RCHY1 (Pirh2) knockout HeLa cell lysate (AB258171)

Allele-2 : 4 bp deletion in exon1

Sanger Sequencing - Human RCHY1 (Pirh2) knockout HeLa cell lysate (AB258171)
  • Sanger seq

Unknown

Sanger Sequencing - Human RCHY1 (Pirh2) knockout HeLa cell lysate (AB258171)

Allele-3 : 1 bp insertion in exon1

Sanger Sequencing - Human RCHY1 (Pirh2) knockout HeLa cell lysate (AB258171)
  • Sanger seq

Unknown

Sanger Sequencing - Human RCHY1 (Pirh2) knockout HeLa cell lysate (AB258171)

Allele-1 : 11 bp deletion in exon1

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 11 bp deletion in exon1 and 1 bp insertion in exon1 and 4 bp deletion in exon1.

Disease

Adenocarcinoma

style
left-column

Complementary Products

Primary Antibodies

AB189247

Anti-Pirh2 antibody [EPR14980]

RabMAb|Recombinant|KO Validated|Trial Size

Immunocytochemistry/ Immunofluorescence - Anti-Pirh2 antibody [EPR14980] (AB189247)

  • 0
  • 0
Proteins & Peptides

AB259412

Recombinant human VEGF 165A protein (Active)

Premium Bioactive

Mass Spectrometry - Recombinant human VEGF 165A protein (Active) (AB259412)

  • 0
  • 0
Proteins & Peptides

AB180317

Recombinant Human POLR1C protein

SDS-PAGE - Recombinant Human POLR1C protein (AB180317)

  • 0
  • 0
Proteins & Peptides

AB288799

Recombinant Human VCAM1 protein (Active)

Premium Bioactive

Biological Activity - Recombinant Human VCAM1 protein (Active) (AB288799)

  • 0
  • 0
style
left-column

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
style
left-column

Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

style
left-column

What's included?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab258171-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human RCHY1 knockout HeLa cell lysate", "number":"AB258171-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HeLa cell lysate", "number":"AB258171-CMP02", "productcode":"" } ] } } }
style
left-column

Properties and storage information

Gene name
RCHY1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
style
left-column

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.
style
left-column

Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

style
left-column

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

style
left-column

Product protocols

style
left-column
style
left-column
style
right-column

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