Anti-Nectin 3/NECTIN3 Antibody Picoband™

Product Name

Anti-Nectin 3/NECTIN3 Antibody Picoband™

View all Nectin-3/PVRL3 Antibodies

SKU/Catalog Number

A09633-3

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-Nectin 3/NECTIN3 Antibody Picoband™ catalog # A09633-3. Tested in ELISA, Flow Cytometry, IHC, WB applications. This antibody reacts with Human.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-Nectin 3/NECTIN3 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A09633-3)

Host

Rabbit

Contents

Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na₂HPO₄, 0.05mg NaN₃.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human Nectin 3/NECTIN3 recombinant protein (Position: E92-N296).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A09633-3 is reactive to NECTIN3 in Human

Applications

A09633-3 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

80 kDa

Calculated molecular weight

61.002kDa

Background of Nectin-3/PVRL3

Nectin-3, also known as poliovirus receptor-related 3, is a 95 kDa type I transmembrane glycoprotein belonging to the Nectin family of Ig superfamily proteins. It is mapped to 3q13.13. This gene encodes a member of the nectin family of proteins, which function as adhesion molecules at adherens junctions. This family member interacts with other nectin-like proteins and with afadin, a filamentous actin-binding protein involved in the regulation of directional motility, cell proliferation and survival. This gene plays a role in ocular development involving the ciliary body. Mutations in this gene are believed to result in congenital ocular defects. Alternative splicing results in multiple transcript variants.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.25-0.5μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human
Flow Cytometry, 1-3μg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5μg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of Nectin 3/NECTIN3 using anti-Nectin 3/NECTIN3 antibody (A09633-3).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human U20S whole cell lysates.
Lane 2: human HEK293 whole cell lysates,
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Nectin 3/NECTIN3 antigen affinity purified polyclonal antibody (Catalog # A09633-3) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Nectin 3/NECTIN3 at approximately 80KD. The expected band size for Nectin 3/NECTIN3 is at 61KD.

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Figure 2. IHC analysis of Nectin 3/NECTIN3 using anti-Nectin 3/NECTIN3 antibody (A09633-3).
Nectin 3/NECTIN3 was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Nectin 3/NECTIN3 Antibody (A09633-3) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

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Figure 3. Flow Cytometry analysis of A431 cells using anti-Nectin 3/NECTIN3 antibody (A09633-3).
Overlay histogram showing A431 cells stained with A09633-3 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Nectin 3/NECTIN3 Antibody (A09633-3,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For NECTIN3 (Source: Uniprot.org, NCBI)

Alternative Names

CD113 antigen; CD113; CDw113FLJ90624; DKFZp566B0846; nectin 3; Nectin3; Nectin-3; poliovirus receptor-related 3; poliovirus receptor-related protein 3; PPR3; PRR3CD113; PVRL3; PVRR3 NECTIN3 CD113, CDW113, NECTIN-3, PPR3, PRR3, PVRL3, PVRR3 nectin cell adhesion molecule 3 nectin-3|nectin 3|poliovirus receptor-related 3|poliovirus receptor-related protein 3

For more info on NECTIN3, check out the NECTIN3 Infographic

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In this infographic, you will see the following information for NECTIN3: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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