Anti-CLDN16/Claudin-16 Antibody Picoband™

Product Name

Anti-CLDN16/Claudin-16 Antibody Picoband™

View all Claudin-16 Antibodies

SKU/Catalog Number

A06659-1

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-CLDN16/Claudin-16 Antibody Picoband™ catalog # A06659-1. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.

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-CLDN16/Claudin-16 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A06659-1)

Host

Rabbit

Contents

Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.01mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

A synthetic peptide corresponding to a sequence at the C-terminus of human Claudin CLDN16/Claudin-16, which shares 95% and 95% amino acid (aa) sequence identity with mouse and rat Claudin CLDN16/Claudin-16, respectively.

*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

A06659-1 is reactive to CLDN16 in Human, Mouse, Rat

Applications

A06659-1 is guaranteed for Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

34 kDa

Calculated molecular weight

33.836kDa

Background of Claudin-16

Claudin-16 is a protein that in humans is encoded by the CLDN16 gene. Tight junctions represent one mode of cell-to-cell adhesion in epithelial or endothelial cell sheets, forming continuous seals around cells and serving as a physical barrier to prevent solutes and water from passing freely through the paracellular space. These junctions are comprised of sets of continuous networking strands in the outwardly facing cytoplasmic leaflet, with complementary grooves in the inwardly facing extracytoplasmic leaflet. The protein encoded by this gene, a member of the claudin family, is an integral membrane protein and a component of tight junction strands. It is found primarily in the kidneys, specifically in the thick ascending limb of Henle, where it acts as either an intercellular pore or ion concentration sensor to regulate the paracellular resorption of magnesium ions. Defects in this gene are a cause of primary hypomagnesemia, which is characterized by massive renal magnesium wasting with hypomagnesemia and hypercalciuria, resulting in nephrocalcinosis and renal failure. This gene and the CLDN1 gene are clustered on chromosome 3q28.

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, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry, 1-3μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of CLDN16/Claudin-16 using anti-CLDN16/Claudin-16 antibody (A06659-1).
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: mouse kidney tissue lysates,
Lane 2: rat NRK 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-CLDN16/Claudin-16 antigen affinity purified polyclonal antibody (Catalog # A06659-1) 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 1:5000 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 CLDN16/Claudin-16 at approximately 34KD. The expected band size for CLDN16/Claudin-16 is at 34KD.

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Figure 2. IF analysis of CLDN16/Claudin-16 using anti-CLDN16/Claudin-16 antibody (A06659-1).
CLDN16/Claudin-16 was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-CLDN16/Claudin-16 Antibody (A06659-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Figure 3. Flow Cytometry analysis of PC-3 cells using anti-CLDN16/Claudin-16 antibody (A06659-1).
Overlay histogram showing PC-3 cells stained with A06659-1 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CLDN16/Claudin-16 Antibody (A06659-1, 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 CLDN16 (Source: Uniprot.org, NCBI)

Alternative Names

claudin 16; Claudin16; Claudin-16; CLDN16; HOMG3; HOMG3paracellin-1; Paracellin-1; PCLN1; PCLN-1; PCLN1hypomagnesemia 3, with hypercalciuria and nephrocalcinosis CLDN16 HOMG3, PCLN1 claudin 16 claudin-16|hypomagnesemia 3, with hypercalciuria and nephrocalcinosis|paracellin-1

For more info on CLDN16, check out the CLDN16 Infographic

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In this infographic, you will see the following information for CLDN16: 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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