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Emi1 Recombinant Rabbit Monoclonal Antibody [JG35-83]

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Specification

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Catalog# ET7108-20

Emi1 Recombinant Rabbit Monoclonal Antibody [JG35-83]

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

Emi1 Recombinant Rabbit Monoclonal Antibody [JG35-83]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human Emi1 aa 128-244 / 447.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P

Molecular Weight

Predicted band size: 50 kDa

Positive Control

SK-OV-3 cell lysate, SiHa cell lysate, LOVO, Siha, human brain tissue, human kidney tissue, human colon carcinoma tissue, mouse brain tissue, mouse hippocampus tissue, rat cerebellum tissue.

Conjugation

unconjugated

Clone Number

JG35-83

RRID

AB_3070820

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

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:500

  • IF-Cell

  • 1:50-1:200

  • IHC-P

  • 1:100-1:1,000

Target

Function

Emi1 (for early mitotic inhibitor) regulates mitosis by inhibiting the anaphase promoting complex/cyclosome (APC). Emi1 is a conserved F box protein containing a zinc binding region essential for APC inhibition. The Emi1 protein functions to promote cyclin A accumulation and S phase entry in somatic cells by inhibiting the APC complex. At the G1-S transition, Emi1 is transcriptionally induced by the E2F transcription factor. Emi1 overexpression accelerates S phase entry and can override a G1 block caused by overexpression of Cdh1 or the E2F-inhibitor p105 retinoblastoma protein (pRb). Depleting cells of Emi1 through RNA interference prevents accumulation of cyclin A and inhibits S phase entry. Emi1 is required to arrest unfertilized eggs at metaphase of meiosis II and may be the long-sought mediator of CSF activity. Human Emi1 is similar to Xenopus laevis Emi1, which inhibits the APC (Cdc20) ubiquitination complex to allow accumulation of cyclin B.

Background References

1. Hsu J Y et al. E2F-dependent accumulation of hEmi1 regulates S phase entry by inhibiting APC(Cdh1). Nat Cell Biol 4:358-366 (2002).

2. Miller J J et al. Emi1 stably binds and inhibits the anaphase-promoting complex/cyclosome as a pseudosubstrate inhibitor. Genes Dev 20:2410-2420 (2006).

Post-translational Modification

Phosphorylation by CDK2 and subsequently by PLK1 triggers degradation during early mitosis through ubiquitin-mediated proteolysis by the SCF ubiquitin ligase complex containing the F-box protein BTRC. This degradation is necessary for the activation of APC in late mitosis and subsequent mitotic progression. Phosphorylated by RPS6KA2; increases and stabilizes interaction with CDC20 (By similarity).; Ubiquitinated by the SCF(BTRC) complex following phosphorylation by PLK1. Undergoes both 'Lys-11' and 'Lys-48'-linked polyubiquitination by APC-FZR1 complex leading to degradation by proteasome during G1 phase. Degraded through the SCF(BTRC) complex; degradation occurs during oocyte maturation, between germinal vesicle breakdown (GVBD) and meiosis I, and is required for the meiosis I-meiosis II transition (By similarity).

Subcellular Location

Cytoplasm. Cytoskeleton. Nucleus.

UNIPROT

SwissProt: Q9UKT4 Human

SwissProt: Q7TSG3 Mouse

SwissProt: B0BNA4 Rat

Synonyms

Early mitotic inhibitor 1 antibody

Emi 1 antibody

EMI1 antibody

F box only protein 5 antibody

F box protein 5 antibody

F box protein Fbx 5 antibody

F box protein Fbx5 antibody

F-box only protein 5 antibody

FBX5 antibody

FBX5_HUMAN antibody

Expand

Images

  • Western blot analysis of Emi1 on different lysates with Rabbit anti-Emi1 antibody (<a href="/products/ET7108-20" style="font-weight: bold;text-decoration: underline;">ET7108-20</a>) at 1/500 dilution.<br /><br />Lane 1: SK-OV-3 cell lysate<br />Lane 2: SiHa cell lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br /><br />Predicted band size: 50 kDa<br />Observed band size: 40-60 kDa<br /><br />Exposure time: Lane 1: 40 seconds; Lane 2: 3 minutes; ECL: K1801;<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/ET7108-20" style="font-weight: bold;text-decoration: underline;">ET7108-20</a>) at 1/500 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 Emi1 on different lysates with Rabbit anti-Emi1 antibody (ET7108-20) at 1/500 dilution.

    Lane 1: SK-OV-3 cell lysate
    Lane 2: SiHa cell lysate

    Lysates/proteins at 15 µg/Lane.

    Predicted band size: 50 kDa
    Observed band size: 40-60 kDa

    Exposure time: Lane 1: 40 seconds; Lane 2: 3 minutes; 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 (ET7108-20) at 1/500 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.

  • Immunocytochemistry analysis of LOVO cells labeling Emi1 with Rabbit anti-Emi1 antibody (<a href="/products/ET7108-20" style="font-weight: bold;text-decoration: underline;">ET7108-20</a>) at 1/50 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Emi1 antibody (<a href="/products/ET7108-20" style="font-weight: bold;text-decoration: underline;">ET7108-20</a>) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃.Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution.Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of LOVO cells labeling Emi1 with Rabbit anti-Emi1 antibody (ET7108-20) at 1/50 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Emi1 antibody (ET7108-20) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃.Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution.Nuclear DNA was labelled in blue with DAPI.

  • Immunocytochemistry analysis of Siha cells labeling Emi1 with Rabbit anti-Emi1 antibody (<a href="/products/ET7108-20" style="font-weight: bold;text-decoration: underline;">ET7108-20</a>) at 1/50 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Emi1 antibody (<a href="/products/ET7108-20" style="font-weight: bold;text-decoration: underline;">ET7108-20</a>) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃.Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of Siha cells labeling Emi1 with Rabbit anti-Emi1 antibody (ET7108-20) at 1/50 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Emi1 antibody (ET7108-20) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃.Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

  • Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-Emi1 antibody (<a href="/products/ET7108-20" style="font-weight: bold;text-decoration: underline;">ET7108-20</a>) at 1/1,000 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/ET7108-20" style="font-weight: bold;text-decoration: underline;">ET7108-20</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 brain tissue with Rabbit anti-Emi1 antibody (ET7108-20) at 1/1,000 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 (ET7108-20) 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-Emi1 antibody (<a href="/products/ET7108-20" style="font-weight: bold;text-decoration: underline;">ET7108-20</a>) at 1/400 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/ET7108-20" style="font-weight: bold;text-decoration: underline;">ET7108-20</a>) at 1/400 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-Emi1 antibody (ET7108-20) at 1/400 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 (ET7108-20) at 1/400 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 colon carcinoma tissue with Rabbit anti-Emi1 antibody (<a href="/products/ET7108-20" style="font-weight: bold;text-decoration: underline;">ET7108-20</a>) at 1/400 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/ET7108-20" style="font-weight: bold;text-decoration: underline;">ET7108-20</a>) at 1/400 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 colon carcinoma tissue with Rabbit anti-Emi1 antibody (ET7108-20) at 1/400 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 (ET7108-20) at 1/400 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-Emi1 antibody (<a href="/products/ET7108-20" style="font-weight: bold;text-decoration: underline;">ET7108-20</a>) at 1/400 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/ET7108-20" style="font-weight: bold;text-decoration: underline;">ET7108-20</a>) at 1/400 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-Emi1 antibody (ET7108-20) at 1/400 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 (ET7108-20) at 1/400 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 hippocampus tissue with Rabbit anti-Emi1 antibody (<a href="/products/ET7108-20" style="font-weight: bold;text-decoration: underline;">ET7108-20</a>) at 1/400 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/ET7108-20" style="font-weight: bold;text-decoration: underline;">ET7108-20</a>) at 1/400 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 hippocampus tissue with Rabbit anti-Emi1 antibody (ET7108-20) at 1/400 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 (ET7108-20) at 1/400 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 cerebellum tissue with Rabbit anti-Emi1 antibody (<a href="/products/ET7108-20" style="font-weight: bold;text-decoration: underline;">ET7108-20</a>) at 1/100 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/ET7108-20" style="font-weight: bold;text-decoration: underline;">ET7108-20</a>) at 1/100 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 cerebellum tissue with Rabbit anti-Emi1 antibody (ET7108-20) at 1/100 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 (ET7108-20) at 1/100 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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