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RNF20 Recombinant Rabbit Monoclonal Antibody [SP00-47]

Usd: 385

Specification

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Catalog# ET1604-4

RNF20 Recombinant Rabbit Monoclonal Antibody [SP00-47]

Application

  • WB

  • FC

  • IF-Cell

Reactivity

  • Human

  • Mouse

Conjugation

  • unconjugated

Overview

Product Name

RNF20 Recombinant Rabbit Monoclonal Antibody [SP00-47]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human RNF20 aa 480-585/975.

Species Reactivity

Human, Mouse

Validated Applications

WB, FC, IF-Cell

Target Molecular Weight

Predicted band size: 114 kDa

Positive Control

HeLa cell lysate, MCF7 cell lysate, C2C12 cell lysate, A549.

Conjugation

unconjugated

Clone Number

SP00-47

RRID

AB_3069697

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 1:50-1:200

  • FC

  • 1:50-1:100

Target

Function

Ubiquitination is an important mechanism through which three classes of enzymes act in concert to target short-lived or abnormal proteins for destruction. The three classes of enzymes involved in ubiquitination are the ubiquitin-activating enzymes (E1s), the ubiquitin-conjugating enzymes (E2s) and the ubiquitin-protein ligases (E3s). RNF20 (ring finger protein 20), also known as BRE1, BRE1A or hBRE1, is a 975 amino acid nuclear protein that belongs to the BRE1 family. As a component of the RNF20/40 complex, RNF20 functions as an E3 ubiquitin-protein ligase that regulates the monoubiquitination and subsequent degradation of select residues on target proteins, such as Histone H2B. RNF20 is required for transcriptional activation of Hox genes and is most likely recruited by p53 to the MDM2 promoter, thereby acting as a transcriptional co-activator. RNF20 contains one zinc finger domain and exists as a homodimer.

Background References

1. Long L et al. The U4/U6 recycling factor SART3 has histone chaperone activity and associates with USP15 to regulate H2B deubiquitination. J Biol Chem 289:8916-30 (2014).

2. Vethantham V et al. Dynamic loss of H2B ubiquitylation without corresponding changes in H3K4 trimethylation during myogenic differentiation. Mol Cell Biol 32:1044-55 (2012).

Sequence Similarity

Belongs to the BRE1 family.

Tissue Specificity

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

Subcellular Location

Nucleus.

UNIPROT

SwissProt: Q5VTR2 Human

SwissProt: Q5DTM8 Mouse

Synonyms

BRE1 A antibody

BRE1 antibody

BRE1 E3 ubiquitin ligase homolog antibody

BRE1-A antibody

BRE1A antibody

BRE1A_HUMAN antibody

E3 ubiquitin-protein ligase BRE1A antibody

hBRE1 antibody

Homolog of S. cerevisiae BRE1 antibody

RING finger protein 20 antibody

Expand

Images

  • Western blot analysis of RNF20 on different lysates with Rabbit anti-RNF20 antibody (<a href="/products/ET1604-4" style="font-weight: bold;text-decoration: underline;">ET1604-4</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: MCF7 cell lysate<br />Lane 3: C2C12 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 114 kDa<br />Observed band size: 114 kDa<br /><br />Exposure time: 1 minute; ECL: K1802;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1604-4" style="font-weight: bold;text-decoration: underline;">ET1604-4</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of RNF20 on different lysates with Rabbit anti-RNF20 antibody (ET1604-4) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: MCF7 cell lysate
    Lane 3: C2C12 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 114 kDa
    Observed band size: 114 kDa

    Exposure time: 1 minute; ECL: K1802;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1604-4) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Flow cytometric analysis of RNF20 was done on A549 cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ET1604-4" style="font-weight: bold;text-decoration: underline;">ET1604-4</a>, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor&reg;488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).

    Flow cytometric analysis of RNF20 was done on A549 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1604-4, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).

  • Immunocytochemistry analysis of A549 cells labeling RNF20 with Rabbit anti-RNF20 antibody (<a href="/products/ET1604-4" style="font-weight: bold;text-decoration: underline;">ET1604-4</a>) at 1/200 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-RNF20 antibody (<a href="/products/ET1604-4" style="font-weight: bold;text-decoration: underline;">ET1604-4</a>) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of A549 cells labeling RNF20 with Rabbit anti-RNF20 antibody (ET1604-4) at 1/200 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-RNF20 antibody (ET1604-4) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"