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UBE4B Recombinant Rabbit Monoclonal Antibody [JE55-76]

Usd: 385

Specification

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Catalog# ET7111-11

UBE4B Recombinant Rabbit Monoclonal Antibody [JE55-76]

Application

  • WB

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

UBE4B Recombinant Rabbit Monoclonal Antibody [JE55-76]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within N-terminal Human UBE4B.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P

Molecular Weight

Predicted band size: 146 kDa

Positive Control

HeLa cell lysate, Raji cell lysate, RAW264.7 cell lysate, PC-12 cell lysate, Mouse testis tissue lysate, Rat testis tissue lysate, human lung carcinoma tissue, human kidney tissue, mouse kidney tissue, rat kidney tissue, rat skeletal muscle tissue.

Conjugation

unconjugated

Clone Number

JE55-76

RRID

AB_3071068

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. 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

  • 1:5,000

  • IHC-P

  • 1:50-1:200

Target

Function

The modification of proteins with ubiquitin is an important cellular mechanism for targeting abnormal or short-lived proteins for degradation. Ubiquitination involves at least three classes of enzymes: ubiquitin-activating enzymes, or E1s, ubiquitin-conjugating enzymes, or E2s, and ubiquitin-protein ligases, or E3s. This gene encodes an additional conjugation factor, E4, which is involved in multiubiquitin chain assembly. This gene is also the strongest candidate in the neuroblastoma tumor suppressor genes. Alternatively spliced transcript variants encoding distinct isoforms have been found for this gene.

Background References

1. Antoniou N. et. al. The Role of E3, E4 Ubiquitin Ligase (UBE4B) in Human Pathologies. Cancers (Basel). 2019 Dec

2. Du C. et. al. UBE4B targets phosphorylated p53 at serines 15 and 392 for degradation. Oncotarget. 2016 Jan

Subcellular Location

Nucleus, Cytoplasm.

UNIPROT

SwissProt: O95155 Human

SwissProt: Q9ES00 Mouse

Entrez Gene: 298652 Rat

Synonyms

4930551I19Rik antibody

4933406G05Rik antibody

AU014668 antibody

D4Bwg0973e antibody

E4 antibody

HDNB1 antibody

Homozygously deleted in neuroblastoma 1 antibody

KIAA0684 antibody

mKIAA0684 antibody

RP23 173D8.1 antibody

Expand

Images

  • Western blot analysis of UBE4B on different lysates with Rabbit anti-UBE4B antibody (<a href="/products/ET7111-11" style="font-weight: bold;text-decoration: underline;">ET7111-11</a>) at 1/5,000 dilution.<br /><br />Lane 1: HeLa cell lysate (10 µg/Lane)<br />Lane 2: Raji cell lysate (10 µg/Lane)<br />Lane 3: RAW264.7 cell lysate (10 µg/Lane)<br />Lane 4: PC-12 cell lysate (10 µg/Lane)<br />Lane 5: Mouse testis tissue lysate (20 µg/Lane)<br />Lane 6: Rat testis tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 146 kDa<br />Observed band size: 146 kDa<br /><br />Exposure time: 6 seconds; ECL: K1801;<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/ET7111-11" style="font-weight: bold;text-decoration: underline;">ET7111-11</a>) at 1/5,000 dilution was used in primary antibody dilution (K1803) 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 UBE4B on different lysates with Rabbit anti-UBE4B antibody (ET7111-11) at 1/5,000 dilution.

    Lane 1: HeLa cell lysate (10 µg/Lane)
    Lane 2: Raji cell lysate (10 µg/Lane)
    Lane 3: RAW264.7 cell lysate (10 µg/Lane)
    Lane 4: PC-12 cell lysate (10 µg/Lane)
    Lane 5: Mouse testis tissue lysate (20 µg/Lane)
    Lane 6: Rat testis tissue lysate (20 µg/Lane)

    Predicted band size: 146 kDa
    Observed band size: 146 kDa

    Exposure time: 6 seconds; ECL: K1801;
    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 (ET7111-11) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockout (KO)</span><br /><br />All lanes: Western blot analysis of UBE4B with anti-UBE4B antibody [JE55-76] (<a href="/products/ET7111-11" style="font-weight: bold;text-decoration: underline;">ET7111-11</a>) at 1:1,000 dilution.<br />Lane 1: Wild-type N2A whole cell lysate (30 µg).<br />Lane 2: UBE4B knockout N2A whole cell lysate (30 µg).<br /><br /><a href="/products/ET7111-11" style="font-weight: bold;text-decoration: underline;">ET7111-11</a> was shown to specifically react with UBE4B in wild-type N2A cells. No band was observed when UBE4B knockdout sample was tested. Wild-type and UBE4B knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET7111-11" style="font-weight: bold;text-decoration: underline;">ET7111-11</a>, 1:1,000) was used in 5% BSA at room temperature for 2 hours.

    ☑ Knockout (KO)

    All lanes: Western blot analysis of UBE4B with anti-UBE4B antibody [JE55-76] (ET7111-11) at 1:1,000 dilution.
    Lane 1: Wild-type N2A whole cell lysate (30 µg).
    Lane 2: UBE4B knockout N2A whole cell lysate (30 µg).

    ET7111-11 was shown to specifically react with UBE4B in wild-type N2A cells. No band was observed when UBE4B knockdout sample was tested. Wild-type and UBE4B knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET7111-11, 1:1,000) was used in 5% BSA at room temperature for 2 hours.

  • Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue with Rabbit anti-UBE4B antibody (<a href="/products/ET7111-11" style="font-weight: bold;text-decoration: underline;">ET7111-11</a>) at 1/200 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/ET7111-11" style="font-weight: bold;text-decoration: underline;">ET7111-11</a>) at 1/200 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 lung carcinoma tissue with Rabbit anti-UBE4B antibody (ET7111-11) at 1/200 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 (ET7111-11) at 1/200 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-UBE4B antibody (<a href="/products/ET7111-11" style="font-weight: bold;text-decoration: underline;">ET7111-11</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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/ET7111-11" style="font-weight: bold;text-decoration: underline;">ET7111-11</a>) at 1/200 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-UBE4B antibody (ET7111-11) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET7111-11) at 1/200 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 kidney tissue with Rabbit anti-UBE4B antibody (<a href="/products/ET7111-11" style="font-weight: bold;text-decoration: underline;">ET7111-11</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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/ET7111-11" style="font-weight: bold;text-decoration: underline;">ET7111-11</a>) at 1/200 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 kidney tissue with Rabbit anti-UBE4B antibody (ET7111-11) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET7111-11) at 1/200 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 rat kidney tissue with Rabbit anti-UBE4B antibody (<a href="/products/ET7111-11" style="font-weight: bold;text-decoration: underline;">ET7111-11</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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/ET7111-11" style="font-weight: bold;text-decoration: underline;">ET7111-11</a>) at 1/200 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 rat kidney tissue with Rabbit anti-UBE4B antibody (ET7111-11) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET7111-11) at 1/200 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 rat skeletal muscle tissue with Rabbit anti-UBE4B antibody (<a href="/products/ET7111-11" style="font-weight: bold;text-decoration: underline;">ET7111-11</a>) at 1/50 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/ET7111-11" style="font-weight: bold;text-decoration: underline;">ET7111-11</a>) at 1/50 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 rat skeletal muscle tissue with Rabbit anti-UBE4B antibody (ET7111-11) at 1/50 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 (ET7111-11) at 1/50 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.

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Citation