RNF20 Recombinant Monoclonal Antibody

Datasheet

Code	CSB-RA227095A0HU
Size	US$210
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Image	Western Blot Positive WB detected in: Hela whole cell lysate, 293 whole cell lysate All lanes: RNF20 antibody at 1:1000 Secondary Goat polyclonal to rabbit IgG at 1/50000 dilution Predicted band size: 114 kDa Observed band size: 114 kDa IHC image of CSB-RA227095A0HU diluted at 1:100 and staining in paraffin-embedded human testis tissue performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4℃ overnight. The primary is detected by a Goat anti-rabbit IgG polymer labeled by HRP and visualized using 0.05% DAB. IHC image of CSB-RA227095A0HU diluted at 1:100 and staining in paraffin-embedded human melanoma cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4℃ overnight. The primary is detected by a Goat anti-rabbit IgG polymer labeled by HRP and visualized using 0.05% DAB. Immunofluorescence staining of Hela Cells with CSB-RA227095A0HU at 1:50, counter-stained with DAPI. The cells were fixed in 4% formaldehyde, permeated by 0.2% TritonX-100, and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4℃. Nuclear DNA was labeled in blue with DAPI. The secondary antibody was FITC-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L).
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Target Background

Product Details

Uniprot No.

Q5VTR2

Target Names

RNF20

Alternative Names

E3 ubiquitin-protein ligase BRE1A (BRE1-A) (hBRE1) (EC 2.3.2.27) (RING finger protein 20) (RING-type E3 ubiquitin transferase BRE1A), RNF20, BRE1A

Species Reactivity

Human

Immunogen

A synthesized peptide derived from human RNF20

Immunogen Species

Homo sapiens (Human)

Conjugate

Non-conjugated

Clonality

Monoclonal

Isotype

Rabbit IgG

Clone No.

2H12

Purification Method

Affinity-chromatography

Concentration

It differs from different batches. Please contact us to confirm it.

Buffer

Rabbit IgG in 10mM phosphate buffered saline , pH 7.4, 150mM sodium chloride, 0.05% BSA, 0.02% sodium azide and 50% glycerol.

Form

Liquid

Tested Applications

ELISA, WB, IHC, IF

Recommended Dilution

Application	Recommended Dilution
WB	1:500-1:5000
IHC	1:50-1:200
IF	1:20-1:200

Protocols

ELISA Protocol
Western Blotting(WB) Protocol
Immunohistochemistry (IHC) Protocol
Immunofluorescence (IF) Protocol

Troubleshooting and FAQs

Antibody FAQs

Storage

Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.

Lead Time

Basically, we can dispatch the products out in 1-3 working days after receiving your orders. Delivery time maybe differs from different purchasing way or location, please kindly consult your local distributors for specific delivery time.

Description

RNF20, also known as BRE1A, functions as an E3 ubiquitin-protein ligase that plays a central role in chromatin regulation through monoubiquitination of histone H2B. This modification serves as a critical epigenetic mark that influences transcriptional elongation, DNA damage response, and genome stability. Researchers investigating chromatin dynamics, transcriptional regulation, and cancer biology frequently target RNF20 due to its involvement in tumor suppression pathways and its emerging connections to developmental processes.

This recombinant monoclonal antibody, generated from clone 2H12 in rabbit host, offers the reproducibility and consistency that demanding experimental workflows require. Because the antibody sequence is defined and production occurs through recombinant expression, researchers can expect minimal lot-to-lot variation, ensuring that results remain comparable across extended studies and collaborative projects. The antibody was raised against a synthetic peptide derived from human RNF20 and purified using affinity chromatography for optimal specificity.

Validation across multiple applications demonstrates this antibody's versatility in the laboratory. Western blot analysis in HeLa and 293 whole cell lysates reveals a clean band at the predicted 114 kDa molecular weight, confirming accurate target recognition at dilutions ranging from 1:500 to 1:5000. Immunohistochemistry studies in paraffin-embedded human testis and melanoma cancer tissues show reliable staining at 1:100 dilution using citrate buffer antigen retrieval. Immunofluorescence analysis in HeLa cells further confirms nuclear localization patterns consistent with RNF20's known chromatin-associated functions.

For cell biology research exploring ubiquitin signaling, epigenetic regulation, or cancer-related pathways, this antibody provides a dependable tool with documented performance across complementary detection methods.

Usage

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

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Target Background

Function

Component of the RNF20/40 E3 ubiquitin-protein ligase complex that mediates monoubiquitination of 'Lys-120' of histone H2B (H2BK120ub1). H2BK120ub1 gives a specific tag for epigenetic transcriptional activation and is also prerequisite for histone H3 'Lys-4' and 'Lys-79' methylation (H3K4me and H3K79me, respectively). It thereby plays a central role inb histone code and gene regulation. The RNF20/40 complex forms a H2B ubiquitin ligase complex in cooperation with the E2 enzyme UBE2A or UBE2B; reports about the cooperation with UBE2E1/UBCH are contradictory. Required for transcriptional activation of Hox genes. Recruited to the MDM2 promoter, probably by being recruited by p53/TP53, and thereby acts as a transcriptional coactivator. Mediates the polyubiquitination of isoform 2 of PA2G4 in cancer cells leading to its proteasome-mediated degradation.

Gene References into Functions

the RNF20/40 complex, a major ubiquitin ligase catalysing histone H2B monoubiquitination, interacts with the motor protein Eg5 during mitosis and participates in spindle assembly. PMID: 27557628
data indicate that RNF20 and PARP1 are synthetic lethal interactors PMID: 28462496
RNF20 depletion stabilizes the ZSCAN4 protein half-life, suggesting that RNF20 negatively regulates ZSCAN4 stability. PMID: 29477841
The effects of RNF20 on mammary tumorigenesis are subtype dependent: In basal-like cancers cells, RNF20 suppresses the NF-kappaB-dependent expression of cytokines, known to contribute to the growth of this tumor subtype. PMID: 28157208
The authors also show that the RING domains of RNF20 and RNF40 can form a stable heterodimer that is active. PMID: 27569044
Manipulation of key H2Bub1 E3 ubiquitin ligases, RNF20, RNF40 and BRCA1, in ovarian cancer cell line models modulated H2Bub1 levels, indicative of the role of these RING finger ligases in monoubiquitination of H2Bub1 in vitro PMID: 27798111
RNF20 and H2Bub1 promotes chronic colonic inflammation and inflammation-associated colorectal cancer in mice and humans, partly by augmenting NF-kappaB activity and attenuating the antitumoral T cell response. PMID: 26854224
It was observed that RE-IIBP induces MEIS1-mediated apoptosis, which was dependent on H2BK120 ubiquitination by RNF20. PMID: 26206755
A primary role for FACT in RNF20 recruitment chromatin remodeling for initiation of homologous recombination repair. PMID: 24357716
The ability of E1A to target hBre1 to simultaneously repress cellular IFN dependent transcription while activating viral transcription PMID: 23785282
model whereby cotranscriptional recruitment of Rnf20 at MLL-fusion target genes leads to amplification of Dot1l-mediated H3K79 methylation, thereby rendering leukemia cells dependent on Rnf20 to maintain their oncogenic transcriptional program PMID: 23412334
We show that Bre1 (human BRE1A/B (RNF20/40) and mouse Bre1a/b (Rnf20/40)) acts as an important suppressor of chromosomal instability PMID: 22354749
our results suggest that RNF20 and RNF40, either via ubiquitylation of H2B or other targets, are coupled to the proliferation of prostate cancer cells. PMID: 22155569
Studies indicate that H2B monoubiquitylation is driven primarily by an E3 ubiquitin ligase composed of the two RING finger proteins RNF20 and RNF40. PMID: 21827756
RNF20, presumably via H2Bub, selectively represses oncogenic genes by interfering with chromatin recruitment of TFIIS, a factor capable of relieving stalled RNA polymerase II. RNF20 inhibits the interaction between TFIIS and the PAF1 complex. PMID: 21596312
RNF20-mediated H2B ubiquitination at DSBs plays a critical role in HRR through chromatin remodeling PMID: 21362548
the observed defects in the radiation response of Bre1a/b-deficient cells PMID: 20738173
RNF20 overexpression leads to elevated histone 2B monoubiquitination, subsequently higher levels of methylation at H3 lysines 4 and 79, and stimulation of homeobox gene expression. PMID: 16307923
The Bre1 protein specifically increases the global level of H2B ubiquitylation at lysine 120 and enhances activator-dependent transcription. PMID: 16337599
identified as one of five genes containing 11 somatic mutations in a panel that included 132 colorectal cancers, then demonstrated that down-regulation of such homologs resulted in chromosomal instability and chromatid cohesion defects in human cells PMID: 18299561
RNF20 suppresses the expression of several proto-oncogenes, which reside preferentially in closed chromatin and are modestly transcribed despite bearing marks usually associated with high transcription rates. PMID: 18832071
hBre1 inhibits Ebp1's tumor suppressive activity through mediating its polyubiquitination and degradation. PMID: 19037095
In humans, the 600 kDa RNF20/40 complex is the E3 ligase. PMID: 16307923
Functional analysis of the yeast counterpart PMID: 12535538

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Subcellular Location

Nucleus.

Protein Families

BRE1 family

Tissue Specificity

Expressed in the normal brain and also in malignant gliomas (at protein level).

Database Links

HGNC: 10062

OMIM: 607699

KEGG: hsa:56254

STRING: 9606.ENSP00000373772

UniGene: Hs.729085

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301-363-4651 (Available 9 a.m. to 5 p.m. CST from Monday to Friday)

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No. 269, Shendun 5th Road, Donghu Hi-Tech Development Area, Hubei Province, 430206, P.R.China

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