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Nectin 2 Recombinant Rabbit Monoclonal Antibody [PSH11-54] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA751416

Nectin 2 Recombinant Rabbit Monoclonal Antibody [PSH11-54] - BSA and Azide free

Application

  • WB

  • IF-Cell

  • FC

Reactivity

  • Human

With BSA and Azide
Conjugation

  • unconjugated

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

Overview

Product Name

Nectin 2 Recombinant Rabbit Monoclonal Antibody [PSH11-54] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human Nectin 2 aa 1-360.

Species Reactivity

Human

Validated Applications

WB, IF-Cell, FC

Molecular Weight

Predicted band size: 58 kDa

Positive Control

MCF7 cell lysate, HT-29 cell lysate, MCF7.

Conjugation

unconjugated

Clone Number

PSH11-54

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

  • 1:2,000

  • IF-Cell

  • 1:100

  • FC

  • 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. Wienke J et al. Integrative analysis of neuroblastoma by single-cell RNA sequencing identifies the NECTIN2-TIGIT axis as a target for immunotherapy. Cancer Cell. 2024 Feb

2. Agorku DJ et al. Colorectal cancer-associated fibroblasts inhibit effector T cells via NECTIN2 signaling. Cancer Lett. 2024 Jul

Subcellular Location

Cell membrane.

UNIPROT

SwissProt: Q92692 Human

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

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of Nectin 2 on different lysates with Rabbit anti-Nectin 2 antibody (<a href="/products/HA751416" style="font-weight: bold;text-decoration: underline;">HA751416</a>) at 1/2,000 dilution.<br /><br />Lane 1: MCF7 cell lysate<br />Lane 2: MOLT-4 cell lysate (negative)<br />Lane 3: HT-29 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 58 kDa<br />Observed band size: 58-75 kDa<br /><br />Exposure time: 25 seconds; ECL: K1802;<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/HA751416" style="font-weight: bold;text-decoration: underline;">HA751416</a>) at 1/2,000 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.

    ☑ Relative expression (RE)

    Western blot analysis of Nectin 2 on different lysates with Rabbit anti-Nectin 2 antibody (HA751416) at 1/2,000 dilution.

    Lane 1: MCF7 cell lysate
    Lane 2: MOLT-4 cell lysate (negative)
    Lane 3: HT-29 cell lysate

    Lysates/proteins at 20 µg/Lane.

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

    Exposure time: 25 seconds; ECL: K1802;

    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 (HA751416) at 1/2,000 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;">☑ Relative expression (RE)</span><br /><br />Immunocytochemistry analysis of MCF7 (positive) and MOLT-4 (negative) labeling Nectin 2 with Rabbit anti-Nectin 2 antibody (<a href="/products/HA751416" style="font-weight: bold;text-decoration: underline;">HA751416</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-Nectin 2 antibody (<a href="/products/HA751416" style="font-weight: bold;text-decoration: underline;">HA751416</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.

    ☑ Relative expression (RE)

    Immunocytochemistry analysis of MCF7 (positive) and MOLT-4 (negative) labeling Nectin 2 with Rabbit anti-Nectin 2 antibody (HA751416) 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-Nectin 2 antibody (HA751416) 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.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Flow cytometric analysis of MOLT-4 (left, negative) and MCF7 (right, positive) cells labeling Nectin 2.<br /><br />Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (<a href="/products/HA751416" style="font-weight: bold;text-decoration: underline;">HA751416</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).

    ☑ Relative expression (RE)

    Flow cytometric analysis of MOLT-4 (left, negative) and MCF7 (right, positive) cells labeling Nectin 2.

    Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA751416, 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).

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation