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Product Info Summary
| SKU: | A04393-5 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-Claudin 3/CLDN3 Antibody Picoband®
SKU/Catalog Number
A04393-5
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Claudin 3/CLDN3 Antibody Picoband® catalog # A04393-5. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, 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-Claudin 3/CLDN3 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A04393-5)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human Claudin 3/CLDN3 recombinant protein (Position: A101-V220).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A04393-5 is reactive to CLDN3 in Human
Observed Molecular Weight
23 kDa
Calculated molecular weight
23.3 kDa
Background of CLDN3
Claudin 3, also known as CLDN3, is a protein which in humans is encoded by the CLDN3 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 intronless gene, a member of the claudin family, is an integral membrane protein and a component of tight junction strands. It is also a low-affinity receptor for Clostridium perfringens enterotoxin, and shares aa sequence similarity with a putative apoptosis-related protein found in rat.
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
A04393-5 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, 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, 0.1-0.25μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: human MCF-7 whole cell, human Caco-2 whole cell
IHC: human renal clear cell carcinoma tissue, human gallbladder adenocarcinoma tissue, human breast invasive ductal cancer tissue, human colonic adenocarcinoma tissue, human ovarian serous adenocarcinoma tissue, human liver cancer tissue
ICC/IF: MCF-7 cell
FCM: MCF-7 cell
Validation Images & Assay Conditions
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Western blot analysis of Claudin 3/CLDN3 using anti-Claudin 3/CLDN3 antibody (A04393-5).
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 30ug of sample under reducing conditions.
Lane 1: human MCF-7 whole cell lysates,
Lane 2: human Caco-2 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-Claudin 3/CLDN3 antigen affinity purified polyclonal antibody (Catalog # A04393-5) at 0.25 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 Claudin 3/CLDN3 at approximately 23KD. The expected band size for Claudin 3/CLDN3 is at 23KD.
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IHC analysis of Claudin 3/CLDN3 using anti-Claudin 3/CLDN3 antibody (A04393-5).
Claudin 3/CLDN3 was detected in paraffin-embedded section of human renal clear cell carcinoma 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 2μg/ml rabbit anti-Claudin 3/CLDN3 Antibody (A04393-5) 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.
Click image to see more details
IHC analysis of Claudin 3/CLDN3 using anti-Claudin 3/CLDN3 antibody (A04393-5).
Claudin 3/CLDN3 was detected in paraffin-embedded section of human gallbladder adenocarcinoma 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 2μg/ml rabbit anti-Claudin 3/CLDN3 Antibody (A04393-5) 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.
Click image to see more details
IHC analysis of Claudin 3/CLDN3 using anti-Claudin 3/CLDN3 antibody (A04393-5).
Claudin 3/CLDN3 was detected in paraffin-embedded section of human breast invasive ductal 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 2μg/ml rabbit anti-Claudin 3/CLDN3 Antibody (A04393-5) 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.
Click image to see more details
IHC analysis of Claudin 3/CLDN3 using anti-Claudin 3/CLDN3 antibody (A04393-5).
Claudin 3/CLDN3 was detected in paraffin-embedded section of human colonic adenocarcinoma 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 2μg/ml rabbit anti-Claudin 3/CLDN3 Antibody (A04393-5) 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.
Click image to see more details
IHC analysis of Claudin 3/CLDN3 using anti-Claudin 3/CLDN3 antibody (A04393-5).
Claudin 3/CLDN3 was detected in paraffin-embedded section of human ovarian serous adenocarcinoma 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 2μg/ml rabbit anti-Claudin 3/CLDN3 Antibody (A04393-5) 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.
Click image to see more details
IHC analysis of Claudin 3/CLDN3 using anti-Claudin 3/CLDN3 antibody (A04393-5).
Claudin 3/CLDN3 was detected in paraffin-embedded section of human liver 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 2μg/ml rabbit anti-Claudin 3/CLDN3 Antibody (A04393-5) 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.
Click image to see more details
IF analysis of Claudin 3/CLDN3 using anti-Claudin 3/CLDN3 antibody (A04393-5).
Claudin 3/CLDN3 was detected in immunocytochemical section of MCF-7 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-Claudin 3/CLDN3 Antibody (A04393-5) 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.
Click image to see more details
Flow Cytometry analysis of MCF-7 cells using anti-Claudin 3/CLDN3 antibody (A04393-5).
Overlay histogram showing MCF-7 cells stained with A04393-5 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-Claudin 3/CLDN3 Antibody (A04393-5, 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-Claudin 3/CLDN3 Antibody Picoband® (A04393-5)
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1 Customer Q&As for Anti-Claudin 3/CLDN3 Antibody Picoband®
Question
Claudin 3 is a cell membrane protein with extracellular and cytoplasmic domains. For the FACS testing data on the datasheet, did you permeabilize the cells? If yes, how (with what for how long)?
Verified customer
Asked: 2022-02-23
Answer
For A04393-5, our lab didn't permeabilize the cells we just fixed the cells for FACS.
Boster Scientific Support
Answered: 2022-02-24
