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Ubiquitin Rabbit Polyclonal Antibody

Usd: 315

Specification

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Catalog# ER31212

Ubiquitin Rabbit Polyclonal Antibody

Application

  • WB

  • IF-Cell

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Ubiquitin Rabbit Polyclonal Antibody

Antibody Type

Rabbit Polyclonal Antibody

Immunogen

Synthetic peptide within human Ubiquitin aa 21-76.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P

Molecular Weight

Predicted band size: 8 kDa

Positive Control

293T cell lysate, HepG2 cell lysate, Hela cell lysate, MCF-7 cell lysate, Hela, HepG2, human tonsil tissue, mouse lung tissue, human kidney tissue.

Conjugation

unconjugated

RRID

AB_3069563

Product Features

Form

Liquid

Concentration

Storage Instructions

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

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Immunogen affinity purified.

Application Dilution

  • WB

  • 1:500-1:1,000

  • IF-Cell

  • 1:100-1:200

  • IHC-P

  • 1:200-1:1,000

Target

Function

Ubiquitin is a conserved polypeptide unit that plays an important role in the ubiquitin-proteasome pathway. Ubiquitin exists either covalently attached to another protein, or free (unanchored). When covalently bound, it is conjugated to target proteins via an isopeptide bond either as a monomer (monoubiquitin), a polymer linked via different Lys residues of the ubiquitin (polyubiquitin chains) or a linear polymer linked via the initiator Met of the ubiquitin (linear polyubiquitin chains). Polyubiquitin chains, when attached to a target protein, have different functions depending on the Lys residue of the ubiquitin that is linked: Lys-6-linked may be involved in DNA repair; Lys-11-linked is involved in ERAD (endoplasmic reticulum-associated degradation) and in cell-cycle regulation; Lys-29-linked is involved in lysosomal degradation; Lys-33-linked is involved in kinase modification; Lys-48-linked is involved in protein degradation via the proteasome; Lys-63-linked is involved in endocytosis, DNA-damage responses as well as in signaling processes leading to activation of the transcription factor NF-kappa-B.

Background References

1. Zhang P et al. Endometrial cancer-associated mutants of SPOP are defective in regulating estrogen receptor-a protein turnover. Cell Death Dis 6:e1687 (2015).

2. Ye X et al. Characterization of PHB1 and its role in mitochondrial maturation and yolk platelet degradation during development of Artemia embryos. PLoS One 9:e109152 (2014).

Sequence Similarity

Belongs to the ubiquitin family.

Post-translational Modification

[Ubiquitin]: Phosphorylated at Ser-65 by PINK1 during mitophagy. Phosphorylated ubiquitin specifically binds and activates parkin (PRKN), triggering mitophagy. Phosphorylation does not affect E1-mediated E2 charging of ubiquitin but affects discharging of E2 enzymes to form polyubiquitin chains. It also affects deubiquitination by deubiquitinase enzymes such as USP30.; [Ubiquitin]: Mono-ADP-ribosylated at the C-terminus by PARP9, a component of the PPAR9-DTX3L complex. ADP-ribosylation requires processing by E1 and E2 enzymes and prevents ubiquitin conjugation to substrates such as histones.

Subcellular Location

Cytoplasm, nucleus, Membrane, Mitochondrion, Mitochondrion outer membrane.

UNIPROT

SwissProt: P0CG48 Human

Synonyms

Epididymis secretory protein Li 50 antibody

FLJ25987 antibody

HEL S 50 antibody

MGC8385 antibody

Polyubiquitin B antibody

RPS 27A antibody

RPS27A antibody

UBA 52 antibody

UBA 80 antibody

UBA52 antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Ubiquitin on different lysates with Rabbit anti-Ubiquitin antibody (<a href="/products/ER31212" style="font-weight: bold;text-decoration: underline;">ER31212</a>) at 1/500 dilution.<br /><br />Lane 1: MCF-7 cell lysate<br />Lane 2: MCF-7 cell lysate treated with 50 µM MG132 for 1.5h<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 8 kDa<br /><br />Exposure time: 2 minutes;<br />15% 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/ER31212" style="font-weight: bold;text-decoration: underline;">ER31212</a>) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:300,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Ubiquitin on different lysates with Rabbit anti-Ubiquitin antibody (ER31212) at 1/500 dilution.

    Lane 1: MCF-7 cell lysate
    Lane 2: MCF-7 cell lysate treated with 50 µM MG132 for 1.5h

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 8 kDa

    Exposure time: 2 minutes;
    15% 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 (ER31212) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of Ubiquitin on different cell lysates using anti-Ubiquitin antibody at 1/1,000 dilution.<br /><span style="font-weight: bold;">Positive control:</span> <br />Lane 1: 293T cell lysate <br />Lane 2: HepG2 cell lysate <br />Lane 3: Hela cell lysate <br />Lane 4: MCF-7 cell lysate

    Western blot analysis of Ubiquitin on different cell lysates using anti-Ubiquitin antibody at 1/1,000 dilution.
    Positive control:
    Lane 1: 293T cell lysate
    Lane 2: HepG2 cell lysate
    Lane 3: Hela cell lysate
    Lane 4: MCF-7 cell lysate

  • Immunocytochemistry analysis of NIH/3T3 cells labeling Ubiquitin with Rabbit anti-Ubiquitin antibody (<a href="/products/ER31212" style="font-weight: bold;text-decoration: underline;">ER31212</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Ubiquitin antibody (<a href="/products/ER31212" style="font-weight: bold;text-decoration: underline;">ER31212</a>) at 1/100 dilution in 1% BSA in PBST 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.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of NIH/3T3 cells labeling Ubiquitin with Rabbit anti-Ubiquitin antibody (ER31212) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Ubiquitin antibody (ER31212) at 1/100 dilution in 1% BSA in PBST 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.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunocytochemistry analysis of PC-12 cells labeling Ubiquitin with Rabbit anti-Ubiquitin antibody (<a href="/products/ER31212" style="font-weight: bold;text-decoration: underline;">ER31212</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Ubiquitin antibody (<a href="/products/ER31212" style="font-weight: bold;text-decoration: underline;">ER31212</a>) at 1/100 dilution in 1% BSA in PBST 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.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of PC-12 cells labeling Ubiquitin with Rabbit anti-Ubiquitin antibody (ER31212) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Ubiquitin antibody (ER31212) at 1/100 dilution in 1% BSA in PBST 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.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • ICC staining Ubiquitin in Hela cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

    ICC staining Ubiquitin in Hela cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

  • ICC staining Ubiquitin in HepG2 cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

    ICC staining Ubiquitin in HepG2 cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

  • ICC staining Ubiquitin in MCF-7 cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

    ICC staining Ubiquitin in MCF-7 cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

  • Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Rabbit anti-Ubiquitin antibody (<a href="/products/ER31212" style="font-weight: bold;text-decoration: underline;">ER31212</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ER31212" style="font-weight: bold;text-decoration: underline;">ER31212</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Rabbit anti-Ubiquitin antibody (ER31212) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER31212) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Ubiquitin antibody (<a href="/products/ER31212" style="font-weight: bold;text-decoration: underline;">ER31212</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ER31212" style="font-weight: bold;text-decoration: underline;">ER31212</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Ubiquitin antibody (ER31212) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER31212) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

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

Citation

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