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Nectin 2 Recombinant Rabbit Monoclonal Antibody [JG39-64]

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Specification

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Catalog# ET7108-95

Nectin 2 Recombinant Rabbit Monoclonal Antibody [JG39-64]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

Nectin 2 Recombinant Rabbit Monoclonal Antibody [JG39-64]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human Nectin 2 aa 370-538 / 538.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, FC

Molecular Weight

Predicted band size: 58 kDa

Positive Control

HT-29 cell lysate, A549 cell lysate, MDA-MB-231 cell lysate, NIH/3T3 cell lysate, Mouse testis tissue lysate, Rat testis tissue lysate, MCF7, human testis tissue, mouse testis tissue, rat testis tissue.

Conjugation

unconjugated

Clone Number

JG39-64

RRID

AB_3070886

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

  • 1:5,000-1:10,000

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:200

  • FC

  • 1:1:1,000

Target

Function

Nectin is a Ca2+-independent homophilic cell adhesion molecule that belongs to the immunoglobulin superfamily. Human nectin is identical to the poliovirus receptor-related protein (PRR) and has been identified as the α-herpesvirus entry mediator. Nectin constitutes a family consisting of at least Nectin 1, 2 and 3; each member has two or three splicing variants. Nectin 2, also designated PRR2/HveB, is ubquitously expressed, with the highest levels of expression in some human neuronal cell lines, fibroblastic cells, keratinocytes and primary activated T lymphocytes. Nectin 2 has two splicing variants, Nectin 2α (short form) and 2δ (long form). Both Nectin 2α and 2δ have a C-terminal conserved motif (E/A-X-Y-V). This motif interacts with the PDZ domain of the F-Actin-binding protein afadin, through which it is linked to the Actin cytoskeleton. The extracellular regions of the splicing variants are identical, but their transmembrane regions and cytoplasmic regions are unique. Nectin 2 mediates the entry of three mutant herpes simplex virus type 1 (HSV-1) strains that do not use HveA as co-receptor, but not wildtype HSV-1 strains. Nectin 2 also mediates the entry of HSV-2 and pseudorabies virus, but not bovine herpes virus type 1. Nectin 2δ is tyrosine phosphorylated in response to cell-cell adhesion.

Background References

1. Warner M S et al. A cell surface protein with herpesvirus entry activity (HveB) confers susceptibility to infection by mutants of herpes simplex virus type 1, herpes simplex virus type 2, and pseudorabies virus. Virology 246:179-189 (1998).

2. Zhu Y et al. Identification of CD112R as a novel checkpoint for human T cells. J Exp Med 213:167-176 (2016).

Sequence Similarity

Belongs to the nectin family.

Tissue Specificity

Ubiquitous.

Subcellular Location

Cell membrane.

UNIPROT

SwissProt: Q92692 Human

SwissProt: P32507 Mouse

SwissProt: Q5FVC5 Rat

Synonyms

CD 112 antibody

CD112 antibody

CD112 antigen antibody

Herpes virus entry mediator B antibody

Herpes virus entry protein B antibody

Herpesvirus entry mediator B antibody

Herpesvirus entry protein B antibody

Hve B antibody

HveB antibody

Nectin-2 antibody

Expand

Images

  • Western blot analysis of Nectin 2 on different lysates with Rabbit anti-Nectin 2 antibody (<a href="/products/ET7108-95" style="font-weight: bold;text-decoration: underline;">ET7108-95</a>) at 1/5,000 dilution.<br /><br />Lane 1: HT-29 cell lysate<br />Lane 2: A549 cell lysate<br />Lane 3: MDA-MB-231 cell lysate<br />Lane 4: NIH/3T3 cell lysate<br />Lane 5: Mouse testis tissue lysate<br />Lane 6: Rat testis tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 58 kDa<br />Observed band size: 75 kDa<br /><br />Exposure time: 16 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/ET7108-95" style="font-weight: bold;text-decoration: underline;">ET7108-95</a>) at 1/5,000 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 Nectin 2 on different lysates with Rabbit anti-Nectin 2 antibody (ET7108-95) at 1/5,000 dilution.

    Lane 1: HT-29 cell lysate
    Lane 2: A549 cell lysate
    Lane 3: MDA-MB-231 cell lysate
    Lane 4: NIH/3T3 cell lysate
    Lane 5: Mouse testis tissue lysate
    Lane 6: Rat testis tissue lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 58 kDa
    Observed band size: 75 kDa

    Exposure time: 16 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 (ET7108-95) at 1/5,000 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.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of Nectin 2 on different lysates with Rabbit anti-Nectin 2 antibody (<a href="/products/ET7108-95" style="font-weight: bold;text-decoration: underline;">ET7108-95</a>) at 1/10,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-Nectin 2 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 58 kDa<br />Observed band size: 70-85 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/ET7108-95" style="font-weight: bold;text-decoration: underline;">ET7108-95</a>) at 1/10,000 dilution was used in 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.

    ☑ Knockdown (KD)

    Western blot analysis of Nectin 2 on different lysates with Rabbit anti-Nectin 2 antibody (ET7108-95) at 1/10,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-Nectin 2 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 58 kDa
    Observed band size: 70-85 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 (ET7108-95) at 1/10,000 dilution was used in 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.

  • Immunocytochemistry analysis of MCF7 cells labeling Nectin 2 with Rabbit anti-Nectin 2 antibody (<a href="/products/ET7108-95" style="font-weight: bold;text-decoration: underline;">ET7108-95</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Nectin 2 antibody (<a href="/products/ET7108-95" style="font-weight: bold;text-decoration: underline;">ET7108-95</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/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</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 MCF7 cells labeling Nectin 2 with Rabbit anti-Nectin 2 antibody (ET7108-95) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Nectin 2 antibody (ET7108-95) 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 (M1305-2, 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.

  • Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-Nectin 2 antibody (<a href="/products/ET7108-95" style="font-weight: bold;text-decoration: underline;">ET7108-95</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/ET7108-95" style="font-weight: bold;text-decoration: underline;">ET7108-95</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 testis tissue with Rabbit anti-Nectin 2 antibody (ET7108-95) 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 (ET7108-95) 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 mouse testis tissue with Rabbit anti-Nectin 2 antibody (<a href="/products/ET7108-95" style="font-weight: bold;text-decoration: underline;">ET7108-95</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/ET7108-95" style="font-weight: bold;text-decoration: underline;">ET7108-95</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 testis tissue with Rabbit anti-Nectin 2 antibody (ET7108-95) 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 (ET7108-95) 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 testis tissue with Rabbit anti-Nectin 2 antibody (<a href="/products/ET7108-95" style="font-weight: bold;text-decoration: underline;">ET7108-95</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/ET7108-95" style="font-weight: bold;text-decoration: underline;">ET7108-95</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 testis tissue with Rabbit anti-Nectin 2 antibody (ET7108-95) 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 (ET7108-95) 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.

  • Flow cytometric analysis of MCF7 cells labeling Nectin 2.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET7108-95" style="font-weight: bold;text-decoration: underline;">ET7108-95</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of MCF7 cells labeling Nectin 2.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET7108-95, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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