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Cullin 3 Recombinant Rabbit Monoclonal Antibody [JE57-80]

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Specification

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Catalog# HA721882

Cullin 3 Recombinant Rabbit Monoclonal Antibody [JE57-80]

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

Cullin 3 Recombinant Rabbit Monoclonal Antibody [JE57-80]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Cullin 3 aa 719-768 / 768.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P

Molecular Weight

Predicted band size: 89 kDa

Positive Control

HeLa cell lysate, Jurkat cell lysate, PC-12 cell lysate, mouse spleen tissue lysate, mouse testis tissue lysate, rat brain tissue lysate, rat spleen tissue lysate, human brain tissue, mouse brain tissue, rat brain tissue, rat hippocampus tissue.

Conjugation

unconjugated

Clone Number

JE57-80

RRID

AB_3096769

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

  • 1:1,000

  • IHC-P

  • 1:500-1:2,000

Target

Function

This gene encodes a member of the cullin protein family. The encoded protein plays a critical role in the polyubiquitination and subsequent degradation of specific protein substrates as the core component and scaffold protein of an E3 ubiquitin ligase complex. Complexes including the encoded protein may also play a role in late endosome maturation. Mutations in this gene are a cause of type 2E pseudohypoaldosteronism. Alternatively spliced transcript variants encoding multiple isoforms have been observed for this gene. Core component of multiple cullin-RING-based BCR (BTB-CUL3-RBX1) E3 ubiquitin-protein ligase complexes which mediate the ubiquitination and subsequent proteasomal degradation of target proteins. BCR complexes and ARIH1 collaborate in tandem to mediate ubiquitination of target proteins. As a scaffold protein may contribute to catalysis through positioning of the substrate and the ubiquitin-conjugating enzyme. The E3 ubiquitin-protein ligase activity of the complex is dependent on the neddylation of the cullin subunit and is inhibited by the association of the deneddylated cullin subunit with TIP120A/CAND1. The functional specificity of the BCR complex depends on the BTB domain-containing protein as the substrate recognition component. BCR(KLHL42) is involved in ubiquitination of KATNA1.

Background References

1. Scott D.C., Rhee D.Y., Duda D.M., Kelsall I.R., Olszewski J.L., Paulo J.A., de Jong A., Ovaa H., Alpi A.F., Harper J.W., Schulman B.A. Two distinct types of E3 ligases work in unison to regulate substrate ubiquitylation. Cell 166:1198-1214 (2016)

2. Jin L., Pahuja K.B., Wickliffe K.E., Gorur A., Baumgartel C., Schekman R., Rape M. Ubiquitin-dependent regulation of COPII coat size and function. Nature 482:495-500 (2012)

Subcellular Location

Nucleus, Cytoplasm.

UNIPROT

SwissProt: Q13618 Human

SwissProt: Q9JLV5 Mouse

SwissProt: B5DF89 Rat

Synonyms

CUL 3 antibody

Cul-3 antibody

cul3 antibody

CUL3_HUMAN antibody

Cullin 3 antibody

Cullin-3 antibody

Cullin3 antibody

KIAA0617 antibody

PHA2E antibody

Images

  • Western blot analysis of Cullin 3 on different lysates with Rabbit anti-Cullin 3 antibody (<a href="/products/HA721882" style="font-weight: bold;text-decoration: underline;">HA721882</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: Jurkat cell lysate (20 µg/Lane)<br />Lane 3: PC-12 cell lysate (20 µg/Lane)<br />Lane 4: Mouse spleen tissue lysate (40 µg/Lane)<br />Lane 5: Mouse testis tissue lysate (40 µg/Lane)<br />Lane 6: Rat brain tissue lysate (40 µg/Lane)<br />Lane 7: Rat spleen tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 89 kDa<br />Observed band size: 89 kDa<br /><br />Exposure time: 3 minutes;<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/HA721882" style="font-weight: bold;text-decoration: underline;">HA721882</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 Cullin 3 on different lysates with Rabbit anti-Cullin 3 antibody (HA721882) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: Jurkat cell lysate (20 µg/Lane)
    Lane 3: PC-12 cell lysate (20 µg/Lane)
    Lane 4: Mouse spleen tissue lysate (40 µg/Lane)
    Lane 5: Mouse testis tissue lysate (40 µg/Lane)
    Lane 6: Rat brain tissue lysate (40 µg/Lane)
    Lane 7: Rat spleen tissue lysate (40 µg/Lane)

    Predicted band size: 89 kDa
    Observed band size: 89 kDa

    Exposure time: 3 minutes;

    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 (HA721882) 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.

  • Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-Cullin 3 antibody (<a href="/products/HA721882" style="font-weight: bold;text-decoration: underline;">HA721882</a>) at 1/500 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/HA721882" style="font-weight: bold;text-decoration: underline;">HA721882</a>) at 1/500 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 brain tissue with Rabbit anti-Cullin 3 antibody (HA721882) at 1/500 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 (HA721882) at 1/500 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 brain tissue with Rabbit anti-Cullin 3 antibody (<a href="/products/HA721882" style="font-weight: bold;text-decoration: underline;">HA721882</a>) at 1/2,000 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/HA721882" style="font-weight: bold;text-decoration: underline;">HA721882</a>) at 1/2,000 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 brain tissue with Rabbit anti-Cullin 3 antibody (HA721882) at 1/2,000 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 (HA721882) at 1/2,000 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 rat brain tissue with Rabbit anti-Cullin 3 antibody (<a href="/products/HA721882" style="font-weight: bold;text-decoration: underline;">HA721882</a>) at 1/2,000 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/HA721882" style="font-weight: bold;text-decoration: underline;">HA721882</a>) at 1/2,000 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 rat brain tissue with Rabbit anti-Cullin 3 antibody (HA721882) at 1/2,000 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 (HA721882) at 1/2,000 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 rat hippocampus tissue with Rabbit anti-Cullin 3 antibody (<a href="/products/HA721882" style="font-weight: bold;text-decoration: underline;">HA721882</a>) at 1/2,000 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/HA721882" style="font-weight: bold;text-decoration: underline;">HA721882</a>) at 1/2,000 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 rat hippocampus tissue with Rabbit anti-Cullin 3 antibody (HA721882) at 1/2,000 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 (HA721882) at 1/2,000 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.

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Citation