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CRBN Recombinant Rabbit Monoclonal Antibody [PSH0-69] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750646

CRBN Recombinant Rabbit Monoclonal Antibody [PSH0-69] - BSA and Azide free

Application

  • WB

  • IHC-P

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

CRBN Recombinant Rabbit Monoclonal Antibody [PSH0-69] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human CRBN aa 1-442 / 442.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IP

Molecular Weight

Predicted band size: 51 kDa

Positive Control

HEK-293 cell lysate, A375 cell lysate, A549 cell lysate, HeLa cell lysate, SH-SY5Y cell lysate, NIH/3T3 cell lysate, MCF7 cell lysate, mouse brain tissue lysate, rat testis tissue lysate, human testis tissue, human brain tissue.

Conjugation

unconjugated

Clone Number

PSH0-69

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IHC-P

  • 1:200

  • IP

  • 1-2μg/sample

Target

Function

Cereblon is a protein that in humans is encoded by the CRBN gene. The gene that encodes the cereblon protein is found on the human chromosome 3, on the short arm at position p26.3 from base pair 3,190,676 to base pair 3,221,394. CRBN orthologs are highly conserved from plants to humans. Cereblon forms an E3 ubiquitin ligase complex with damaged DNA binding protein 1 (DDB1), Cullin-4A (CUL4A), and regulator of cullins 1 (ROC1). This complex ubiquitinates a number of other proteins. Through a mechanism which has not been completely elucidated, this ubiquitination results in reduced levels of fibroblast growth factor 8 (FGF8) and fibroblast growth factor 10 (FGF10). FGF8 in turn regulates a number of developmental processes, such as limb and auditory vesicle formation. The net result is that this ubiquitin ligase complex is important for limb outgrowth in embryos. In the absence of cereblon, DDB1 forms a complex with DDB2 that functions as a DNA damage-binding protein. Furthermore, cereblon and DDB2 bind to DDB1 in a competitive manner.

Background References

1. Wang C et al. Developments of CRBN-based PROTACs as potential therapeutic agents. Eur J Med Chem. 2021 Dec

2. Yamamoto J et al. Discovery of CRBN as a target of thalidomide: a breakthrough for progress in the development of protein degraders. Chem Soc Rev. 2022 Aug

Subcellular Location

Cytoplasm, Nucleus, Membrane.

UNIPROT

SwissProt: Q96SW2 Human

SwissProt: Q8C7D2 Mouse

SwissProt: Q56AP7 Rat

Synonyms

2610203G15Rik antibody

2900045O07Rik antibody

AF229032 antibody

AW108261 antibody

Cereblon antibody

Crbn antibody

CRBN_HUMAN antibody

DKFZp781K0715 antibody

MGC27358 antibody

MRT2A antibody

Expand

Images

  • Western blot analysis of CRBN on different lysates with Rabbit anti-CRBN antibody (<a href="/products/HA750646" style="font-weight: bold;text-decoration: underline;">HA750646</a>) at 1/1,000 dilution.<br /><br />Lane 1: HEK-293 cell lysate<br />Lane 2: A375 cell lysate<br />Lane 3: A549 cell lysate<br />Lane 4: HeLa cell lysate<br />Lane 5: SH-SY5Y cell lysate<br />Lane 6: NIH/3T3 cell lysate<br />Lane 7: MCF7 cell lysate<br />Lane 8: Mouse brain tissue lysate<br />Lane 9: Rat testis tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 51 kDa<br />Observed band size: 51 kDa<br /><br />Exposure time: 40 seconds;<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/HA750646" style="font-weight: bold;text-decoration: underline;">HA750646</a>) at 1/1,000 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 1:100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of CRBN on different lysates with Rabbit anti-CRBN antibody (HA750646) at 1/1,000 dilution.

    Lane 1: HEK-293 cell lysate
    Lane 2: A375 cell lysate
    Lane 3: A549 cell lysate
    Lane 4: HeLa cell lysate
    Lane 5: SH-SY5Y cell lysate
    Lane 6: NIH/3T3 cell lysate
    Lane 7: MCF7 cell lysate
    Lane 8: Mouse brain tissue lysate
    Lane 9: Rat testis tissue lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 51 kDa
    Observed band size: 51 kDa

    Exposure time: 40 seconds;

    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 (HA750646) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-CRBN antibody (<a href="/products/HA750646" style="font-weight: bold;text-decoration: underline;">HA750646</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/HA750646" style="font-weight: bold;text-decoration: underline;">HA750646</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 testis tissue with Rabbit anti-CRBN antibody (HA750646) 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 (HA750646) 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 brain tissue with Rabbit anti-CRBN antibody (<a href="/products/HA750646" style="font-weight: bold;text-decoration: underline;">HA750646</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/HA750646" style="font-weight: bold;text-decoration: underline;">HA750646</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 brain tissue with Rabbit anti-CRBN antibody (HA750646) 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 (HA750646) 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.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of CRBN on different lysates with Rabbit anti-CRBN antibody (<a href="/products/HA750646" style="font-weight: bold;text-decoration: underline;">HA750646</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa-si NT cell lysate<br />Lane 2: HeLa-si CRBN cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 51 kDa<br />Observed band size: 51 kDa<br /><br />Exposure time: 2 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/HA750646" style="font-weight: bold;text-decoration: underline;">HA750646</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.

    ☑ Knockdown (KD)

    Western blot analysis of CRBN on different lysates with Rabbit anti-CRBN antibody (HA750646) at 1/1,000 dilution.

    Lane 1: HeLa-si NT cell lysate
    Lane 2: HeLa-si CRBN cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 51 kDa
    Observed band size: 51 kDa

    Exposure time: 2 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 (HA750646) 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.

  • CRBN was immunoprecipitated in 0.2mg HeLa cell lysate with <a href="/products/HA750646" style="font-weight: bold;text-decoration: underline;">HA750646</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA750646" style="font-weight: bold;text-decoration: underline;">HA750646</a> at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: HeLa cell lysate (input)<br />Lane 2: <a href="/products/HA750646" style="font-weight: bold;text-decoration: underline;">HA750646</a> IP in HeLa cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA750646" style="font-weight: bold;text-decoration: underline;">HA750646</a> in HeLa cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 1 minute 10 seconds; ECL: <a href="/products/K1802" style="font-weight: bold;text-decoration: underline;">K1802</a>

    CRBN was immunoprecipitated in 0.2mg HeLa cell lysate with HA750646 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA750646 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: HeLa cell lysate (input)
    Lane 2: HA750646 IP in HeLa cell lysate
    Lane 3: Rabbit IgG instead of HA750646 in HeLa cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 1 minute 10 seconds; ECL: K1802

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