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Product Info Summary
| SKU: | A09633-3 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-Nectin 3/NECTIN3 Antibody Picoband®
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. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
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).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A09633-3 is reactive to NECTIN3 in Human
Observed Molecular Weight
80 kDa
Calculated molecular weight
61.0 kDa
Background of NECTIN3
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.
Application & Images
Applications
A09633-3 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee
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 (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: human U20S whole cellhuman HEK293 whole cell,
IHC: human rectal cancer tissue
FCM: A431 cell
Validation Images & Assay Conditions
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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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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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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). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. 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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-Nectin 3/NECTIN3 Antibody Picoband® (A09633-3)
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