Recombinant Anti-p53R2 antibody [EPR8816] KO Tested (ab154194)

Product name

Anti-p53R2 antibody [EPR8816]
See all p53R2 primary antibodies
Description

Rabbit monoclonal [EPR8816] to p53R2
Host species

Rabbit
Tested applications

Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)more details
Species reactivity

Reacts with: Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: MCF7, HCT116, Human skeletal muscle, and SW480 lysates. IHC-P: human breast carcinoma tissue. ICC/IF: HeLa cells. Flow Cyt (intra): MCF7 cells. IP: MCF7 cells.
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.
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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at -20ºC.
Storage buffer

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

Protein A purified
Clonality

Monoclonal
Clone number

EPR8816
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
- Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
- Human RRM2B (p53R2) knockout HeLa cell line (ab261769)
- Human RRM2B (p53R2) knockout HCT116 cell line (ab266897)
KO cell lysates
- Human RRM2B (p53R2) knockout HeLa cell lysate (ab257215)
- Human RRM2B (p53R2) knockout HCT116 cell lysate (ab257216)
Positive Controls
- SW480 whole cell lysate (ab3957)
Recombinant Protein
- Recombinant Human p53R2 protein (ab153320)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab154194 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
WB		1/1000 - 1/10000. Predicted molecular weight: 40 kDa.
IHC-P		1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. For unpurified use at 1/50 - 1/100
ICC/IF		1/50.
IP		1/10 - 1/100.
Flow Cyt (Intra)		1/10 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Notes
WB 1/1000 - 1/10000. Predicted molecular weight: 40 kDa.
IHC-P 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. For unpurified use at 1/50 - 1/100
ICC/IF 1/50.
IP 1/10 - 1/100.
Flow Cyt (Intra) 1/10 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Function

Plays a pivotal role in cell survival by repairing damaged DNA in a p53/TP53-dependent manner. Supplies deoxyribonucleotides for DNA repair in cells arrested at G1 or G2. Contains an iron-tyrosyl free radical center required for catalysis. Forms an active ribonucleotide reductase (RNR) complex with RRM1 which is expressed both in resting and proliferating cells in response to DNA damage.
Tissue specificity

Widely expressed at a high level in skeletal muscle and at a weak level in thymus. Expressed in epithelial dysplasias and squamous cell carcinoma.
Pathway

Genetic information processing; DNA replication.
Involvement in disease

Defects in RRM2B are the cause of mitochondrial DNA depletion syndrome type 8A (MTDPS8A) [MIM:612075]. A disorder due to mitochondrial dysfunction characterized by various combinations of neonatal hypotonia, neurological deterioration, respiratory distress, lactic acidosis, and renal tubulopathy.
Defects in RRM2B are the cause of mitochondrial DNA depletion syndrome type 8B (MTDPS8B) [MIM:612075]. A disease due to mitochondrial dysfunction and characterized by ophthalmoplegia, ptosis, gastrointestinal dysmotility, cachexia, peripheral neuropathy.
Defects in RRM2B are the cause of progressive external ophthalmoplegia with mitochondrial DNA deletions autosomal dominant type 5 (PEOA5) [MIM:613077]. A disorder characterized by progressive weakness of ocular muscles and levator muscle of the upper eyelid. In a minority of cases, it is associated with skeletal myopathy, which predominantly involves axial or proximal muscles and which causes abnormal fatigability and even permanent muscle weakness. Ragged-red fibers and atrophy are found on muscle biopsy. A large proportion of chronic ophthalmoplegias are associated with other symptoms, leading to a multisystemic pattern of this disease. Additional symptoms are variable, and may include cataracts, hearing loss, sensory axonal neuropathy, ataxia, depression, hypogonadism, and parkinsonism.
Sequence similarities

Belongs to the ribonucleoside diphosphate reductase small chain family.
Cellular localization

Cytoplasm. Nucleus. Translocates from cytoplasm to nucleus in response to DNA damage.
Target information above from: UniProt accession Q7LG56 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 50484 Human
- Omim: 604712 Human
- SwissProt: Q7LG56 Human
- Unigene: 512592 Human
Alternative names
- DKFZp686M05248 antibody
- MGC102856 antibody
- MGC42116 antibody
- MTDPS8A antibody
- MTDPS8B antibody
- p53 inducible ribonucleotide reductase small subunit 2 homolog antibody
- p53 inducible ribonucleotide reductase small subunit 2 like protein antibody
- P53 inducible ribonucleotide reductase small subunit 2 short form beta antibody
- p53 R2 antibody
- p53-inducible ribonucleotide reductase small subunit 2-like protein antibody
- p53R2 antibody
- Ribonucleoside diphosphate reductase M2 subunit B antibody
- Ribonucleoside-diphosphate reductase subunit M2 B antibody
- Ribonucleotide reductase M2 B (TP53 inducible) antibody
- Ribonucleotide reductase M2 B antibody
- Ribonucleotide reductase small subunit like 2 p53 inducible antibody
- RIR2B_HUMAN antibody
- RRM 2B antibody
- RRM2B antibody
- TP53 inducible ribonucleotide reductase M2 B antibody
- TP53-inducible ribonucleotide reductase M2 B antibody
see all

Western blot - Anti-p53R2 antibody [EPR8816] (ab154194)

All lanes : Anti-p53R2 antibody [EPR8816] (ab154194) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : RRM2B knockout HeLa cell lysate

Lysates/proteins at 40 µg per lane.

Performed under reducing conditions.

Predicted band size: 40 kDa
Observed band size: 40 kDa

Lanes 1- 2: Merged signal (red and green). Green - ab154194 observed at 40 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

ab154194 was shown to react with p53R2 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261769 (knockout cell lysate ab257215) was used. Wild-type HeLa and RRM2B knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab154194 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 1000 dilution and a 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-p53R2 antibody [EPR8816] (ab154194)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling p53R2 with purified ab154194 at 1/50 dilution (2.2 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53R2 antibody [EPR8816] (ab154194)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling p53R2 with purified ab154194 at 1/500 dilution (0.22 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody.

Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Flow Cytometry (Intracellular) - Anti-p53R2 antibody [EPR8816] (ab154194)

Intracellular Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling p53R2 with purified ab154194 at 1/20 dilution (10µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
Western blot - Anti-p53R2 antibody [EPR8816] (ab154194)

All lanes : Anti-p53R2 antibody [EPR8816] (ab154194) at 1/1000 dilution

Lane 1 : Wild-type HCT116 cell lysate
Lane 2 : RRM2B knockout HCT116 cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 40 kDa
Observed band size: 40 kDa

Lanes 1- 2: Merged signal (red and green). Green - ab154194 observed at 40 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

ab154194 was shown to react with p53R2 in wild-type HCT116 cells in western blot. Loss of signal was observed when knockout cell line ab266897 (knockout cell lysate ab257216) was used. Wild-Type HCT116 and RRM2B knockout HCT116 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab154194 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 1000 dilution and a 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.
Western blot - Anti-p53R2 antibody [EPR8816] (ab154194)

All lanes : Anti-p53R2 antibody [EPR8816] (ab154194) at 1/1000 dilution (Purified)

Lane 1 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : Human skeletal muscle lysates
Lane 3 : SW480 (Human colorectal adenocarcinoma epithelial cell) whole cell lysates

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 40 kDa
Observed band size: 41 kDa why is the actual band size different from the predicted?
Western blot - Anti-p53R2 antibody [EPR8816] (ab154194)

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: p53R2 knockout HAP1 cell lysate (20 µg)
Lane 3: MCF7 cell lysate (20 µg)
Lane 4: SW480 cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab154194 observed at 40 kDa. Red - loading control, ab18058, observed at 124 kDa.

Unpurified ab154194 was shown to recognize p53R2 when p53R2 knockout samples were used, along with additional cross-reactive bands. Wild-type and p53R2 knockout samples were subjected to SDS-PAGE. ab154194 and ab18058 (loading control to Vinculin) were both diluted 1/1000 and incubated overnight at 4°C. 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 1/10000 dilution for 1 h at room temperature before imaging.
Western blot - Anti-p53R2 antibody [EPR8816] (ab154194)

All lanes : Anti-p53R2 antibody [EPR8816] (ab154194) at 1/1000 dilution ((unpurified))

Lane 1 : Human fetal muscle lysate
Lane 2 : MCF7 cell lysate
Lane 3 : SW480 cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 40 kDa
Immunoprecipitation - Anti-p53R2 antibody [EPR8816] (ab154194)

ab154194 (purified ) at 1/20 dilution (0.5ug) immunoprecipitating p53R2 in MCF7 whole cell lysate.

Lane 1 (input): MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10ug
Lane 2 (+): ab154194 & MCF7 whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab154194 in MCF7 whole cell lysate

For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.

Blocking and diluting buffer: 5% NFDM/TBST.
Immunocytochemistry/ Immunofluorescence - Anti-p53R2 antibody [EPR8816] (ab154194)

Immunofluorescent staining of MCF7 cells labeling p53R2 with unpurified ab154194 at 1/250 dilution.
Flow Cytometry (Intracellular) - Anti-p53R2 antibody [EPR8816] (ab154194)

Intracellular flow cytometric analysis of permeabilized MCF7 cells labeling p53R2 with unpurifiedab154194 at 1/10 dilution (red) or a rabbit IgG negative control antibody (green).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53R2 antibody [EPR8816] (ab154194)

Immunohistochemical analysis of paraffin-embedded Human ovarian carcinoma tissue labeling p53R2 with unpurified ab154194 at 1/50 dilution.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53R2 antibody [EPR8816] (ab154194)

Immunohistochemical analysis of paraffin embedded Human urinary bladder transitional carcinoma tissue using unpurified ab154194 showing +ve staining.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53R2 antibody [EPR8816] (ab154194)

Immunohistochemical analysis of paraffin embedded Human melanoma tissue using unpurified ab154194 showing +ve staining.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53R2 antibody [EPR8816] (ab154194)

Immunohistochemical analysis of paraffin embedded Human normal kidney tissue unpurified ab154194 showing +ve staining.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53R2 antibody [EPR8816] (ab154194)

Immunohistochemical analysis of paraffin embedded Human normal colon tissue using unpurified ab154194 showing +ve staining.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53R2 antibody [EPR8816] (ab154194)

Immunohistochemical analysis of paraffin embedded Human normal brain tissue using unpurified ab154194 showing +ve staining.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Anti-p53R2 antibody [EPR8816] (ab154194)

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

Click here to view the general protocols

SDS download

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Publishing research using ab154194? Please let us know so that we can cite the reference in this datasheet.

ab154194 has been referenced in 6 publications.

Li Y et al. Antitumor Mechanism of Hydroxycamptothecin via the Metabolic Perturbation of Ribonucleotide and Deoxyribonucleotide in Human Colorectal Carcinoma Cells. Molecules 26:N/A (2021). PubMed: 34443490
Wright GEB et al. Gene expression profiles complement the analysis of genomic modifiers of the clinical onset of Huntington disease. Hum Mol Genet 29:2788-2802 (2020). PubMed: 32898862
Hu CM et al. High Glucose Triggers Nucleotide Imbalance through O-GlcNAcylation of Key Enzymes and Induces KRAS Mutation in Pancreatic Cells. Cell Metab 29:1334-1349.e10 (2019). PubMed: 30853214
Chen J et al. p53R2 as a novel prognostic biomarker in nasopharyngeal carcinoma. BMC Cancer 17:846 (2017). PubMed: 29237424
Jiang C et al. p53R2 overexpression in cervical cancer promotes AKT signaling and EMT, and is correlated with tumor progression, metastasis and poor prognosis. Cell Cycle 16:1673-1682 (2017). PubMed: 28841361
Guo JR et al. Effect of Phyllanthus amarus Extract on 5-Fluorouracil-Induced Perturbations in Ribonucleotide and Deoxyribonucleotide Pools in HepG2 Cell Line. Molecules 21:N/A (2016). PubMed: 27657029

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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

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Pathway

Involvement in disease

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (6)

Customer reviews and Q&As