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14-3-3 epsilon Recombinant Rabbit Monoclonal Antibody [JJ08-40]

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Specification

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Catalog# ET1701-1

14-3-3 epsilon Recombinant Rabbit Monoclonal Antibody [JJ08-40]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

  • Green monkey

Conjugation

  • unconjugated

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

Overview

Product Name

14-3-3 epsilon Recombinant Rabbit Monoclonal Antibody [JJ08-40]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within human 14-3-3 epsilon aa 51-100.

Species Reactivity

Human, Mouse, Rat, Green monkey

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P

Molecular Weight

Predicted band size: 29 kDa

Positive Control

PC-12 cell lysate, COS-1 cell lysate, Mouse brain tissue lysate, Mouse kidney tissue lysate, Rat brain tissue lysate, Rat liver tissue lysate, 293T cell lysate, SH-SY5Y cell lysate, PC-12, Hela, NIH/3T3, human brain tissue, mouse brain tissue, mouse kidney tissue, rat brain tissue, rat liver tissue.

Conjugation

unconjugated

Clone Number

JJ08-40

RRID

AB_3070176

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

  • 1:5,000

  • IF-Cell

  • 1:50-1:200

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:200

Target

Function

14-3-3 protein epsilon is a protein that in humans is encoded by the YWHAE gene. This gene product belongs to the 14-3-3 family of proteins which mediate signal transduction by binding to phosphoserine-containing proteins. This highly conserved protein family is found in both plants and mammals, and this protein is 100% identical to the mouse ortholog. It interacts with CDC25 phosphatases, RAF1 and IRS1 proteins, suggesting its role in diverse biochemical activities related to signal transduction, such as cell division and regulation of insulin sensitivity. It has also been implicated in the pathogenesis of small cell lung cancer, and microdeletions associated with Miller–Dieker syndrome.

Background References

1. Zhu Y et al. Neuroprotective effect of TAT-14-3-3e fusion protein against cerebral ischemia/reperfusion injury in rats. PLoS One 9:e93334 (2014).

2. Hoyo NL et al. Functional EF-hands in neuronal calcium sensor GCAP2 determine its phosphorylation state and subcellular distribution in vivo, and are essential for photoreceptor cell integrity. PLoS Genet 10:e1004480 (2014).

Sequence Similarity

Belongs to the 14-3-3 family.

Subcellular Location

Nucleus, Cytoplasm, Melanosome.

UNIPROT

SwissProt: P62258 Human

SwissProt: P62259 Mouse

SwissProt: P62260 Rat

Synonyms

14 3 3 E antibody

14 3 3 epsilon antibody

14 3 3E antibody

14-3-3 protein epsilon antibody

14-3-3E antibody

1433E_HUMAN antibody

Epididymis luminal protein 2 antibody

FLJ45465 antibody

FLJ53559 antibody

HEL2 antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of 14-3-3 epsilon on different lysates with Rabbit anti-14-3-3 epsilon antibody (<a href="/products/ET1701-1" style="font-weight: bold;text-decoration: underline;">ET1701-1</a>) at 1/1,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-14-3-3 epsilon KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 29 kDa<br />Observed band size: 29 kDa<br /><br />Exposure time: 20 seconds; 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/ET1701-1" style="font-weight: bold;text-decoration: underline;">ET1701-1</a>) at 1/1,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) 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 14-3-3 epsilon on different lysates with Rabbit anti-14-3-3 epsilon antibody (ET1701-1) at 1/1,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-14-3-3 epsilon KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 29 kDa
    Observed band size: 29 kDa

    Exposure time: 20 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 (ET1701-1) at 1/1,000 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.

  • Western blot analysis of 14-3-3 epsilon on different lysates with Rabbit anti-14-3-3 epsilon antibody (<a href="/products/ET1701-1" style="font-weight: bold;text-decoration: underline;">ET1701-1</a>) at 1/5,000 dilution.<br /><br />Lane 1: PC-12 cell lysate (20 µg/Lane)<br />Lane 2: COS-1 cell lysate (20 µg/Lane)<br />Lane 3: Mouse brain tissue lysate (40 µg/Lane)<br />Lane 4: Mouse kidney tissue lysate (40 µg/Lane)<br />Lane 5: Rat brain tissue lysate (40 µg/Lane)<br />Lane 6: Rat liver tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 29 kDa<br />Observed band size: 29 kDa<br /><br />Exposure time: 10 seconds; 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/ET1701-1" style="font-weight: bold;text-decoration: underline;">ET1701-1</a>) at 1/5,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) 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 14-3-3 epsilon on different lysates with Rabbit anti-14-3-3 epsilon antibody (ET1701-1) at 1/5,000 dilution.

    Lane 1: PC-12 cell lysate (20 µg/Lane)
    Lane 2: COS-1 cell lysate (20 µg/Lane)
    Lane 3: Mouse brain tissue lysate (40 µg/Lane)
    Lane 4: Mouse kidney tissue lysate (40 µg/Lane)
    Lane 5: Rat brain tissue lysate (40 µg/Lane)
    Lane 6: Rat liver tissue lysate (40 µg/Lane)

    Predicted band size: 29 kDa
    Observed band size: 29 kDa

    Exposure time: 10 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 (ET1701-1) 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.

  • Western blot analysis of 14-3-3 epsilon on different lysates with Rabbit anti-14-3-3 epsilon antibody (<a href="/products/ET1701-1" style="font-weight: bold;text-decoration: underline;">ET1701-1</a>) at 1/500 dilution.<br /><br />Lane 1: 293T cell lysate<br />Lane 2: SH-SY5Y cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 29 kDa<br />Observed band size: 29 kDa<br /><br />Exposure time: 30 seconds;<br /><br />12% 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/ET1701-1" style="font-weight: bold;text-decoration: underline;">ET1701-1</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.

    Western blot analysis of 14-3-3 epsilon on different lysates with Rabbit anti-14-3-3 epsilon antibody (ET1701-1) at 1/500 dilution.

    Lane 1: 293T cell lysate
    Lane 2: SH-SY5Y cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 29 kDa
    Observed band size: 29 kDa

    Exposure time: 30 seconds;

    12% 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 (ET1701-1) 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.

  • Immunocytochemistry analysis of PC-12 cells labeling 14-3-3 epsilon with Rabbit anti-14-3-3 epsilon antibody (<a href="/products/ET1701-1" style="font-weight: bold;text-decoration: underline;">ET1701-1</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-14-3-3 epsilon antibody (<a href="/products/ET1701-1" style="font-weight: bold;text-decoration: underline;">ET1701-1</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/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</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 14-3-3 epsilon with Rabbit anti-14-3-3 epsilon antibody (ET1701-1) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-14-3-3 epsilon antibody (ET1701-1) 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 (HA601187, 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 of 14-3-3 epsilon in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1701-1" style="font-weight: bold;text-decoration: underline;">ET1701-1</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of 14-3-3 epsilon in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-1, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • ICC staining of 14-3-3 epsilon in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1701-1" style="font-weight: bold;text-decoration: underline;">ET1701-1</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of 14-3-3 epsilon in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-1, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-14-3-3 epsilon antibody (<a href="/products/ET1701-1" style="font-weight: bold;text-decoration: underline;">ET1701-1</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/ET1701-1" style="font-weight: bold;text-decoration: underline;">ET1701-1</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-14-3-3 epsilon antibody (ET1701-1) 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 (ET1701-1) 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 brain tissue with Rabbit anti-14-3-3 epsilon antibody (<a href="/products/ET1701-1" style="font-weight: bold;text-decoration: underline;">ET1701-1</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/ET1701-1" style="font-weight: bold;text-decoration: underline;">ET1701-1</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 brain tissue with Rabbit anti-14-3-3 epsilon antibody (ET1701-1) 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 (ET1701-1) 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-14-3-3 epsilon antibody (<a href="/products/ET1701-1" style="font-weight: bold;text-decoration: underline;">ET1701-1</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/ET1701-1" style="font-weight: bold;text-decoration: underline;">ET1701-1</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-14-3-3 epsilon antibody (ET1701-1) 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 (ET1701-1) 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 brain tissue with Rabbit anti-14-3-3 epsilon antibody (<a href="/products/ET1701-1" style="font-weight: bold;text-decoration: underline;">ET1701-1</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/ET1701-1" style="font-weight: bold;text-decoration: underline;">ET1701-1</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 brain tissue with Rabbit anti-14-3-3 epsilon antibody (ET1701-1) 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 (ET1701-1) 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 liver tissue with Rabbit anti-14-3-3 epsilon antibody (<a href="/products/ET1701-1" style="font-weight: bold;text-decoration: underline;">ET1701-1</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/ET1701-1" style="font-weight: bold;text-decoration: underline;">ET1701-1</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 liver tissue with Rabbit anti-14-3-3 epsilon antibody (ET1701-1) 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 (ET1701-1) 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.

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