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Product Info Summary
| SKU: | PA1972 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-Zonula occludens protein 3/TJP3 Antibody Picoband®
SKU/Catalog Number
PA1972
BA2862-2 is an alternative SKU for this antibody, used in previous lots.
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Zonula occludens protein 3/TJP3 Antibody catalog # PA1972. Tested in Flow Cytometry, ICC/IF, 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-Zonula occludens protein 3/TJP3 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1972)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence at the C-terminus of human Zonula occludens protein 3.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PA1972 is reactive to TJP3 in Human
Observed Molecular Weight
130-140 kDa
Calculated molecular weight
101.4 kDa
Background of TJP3
TJP3 (TIGHT JUNCTION PROTEIN 3), also called ZO3, is a protein that in humans is encoded by the TJP3 gene. TJP3 is a member of the family of membrane-associated guanylate kinase-like proteins (MAGUK) that associate with intracellular junctions (Itoh et al., 1999). Haskins et al. (1998) showed that recombinant Tjp3 binds in vitro translated Tjp1 and binds directly to the cytoplasmic tail of occludin but no binding was detected between Tjp3 and Tjp2. Roh et al. (2002) determined that Tjp3 interacted with PATJ (INADL) and that both proteins colocalized to tight junctions in vivo. In vitro studies showed that the sixth and eighth PDZ domains of PATJ mediated interaction with the Tjp3 C-terminal domain and with Cldn1, respectively.
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
PA1972 is guaranteed for 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, 0.1-0.5μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry(Fixed), 1-3 μg/1x106 cells, Human
Positive Control
WB: human Caco-2 whole cell, human MCF-7 whole cell, human T-47D whole cell
IHC: human fallopian tube tissue
ICC/IF: CACO-2 cell
FCM: CACO-2 cell
Validation Images & Assay Conditions
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Western blot analysis of TJP3 using anti-TJP3 antibody (PA1972).
Electrophoresis was performed on a 8% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Caco-2 whole cell lysates,
Lane 2: human MCF-7 whole cell lysates,
Lane 3: human T-47D whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-TJP3 antigen affinity purified polyclonal antibody (PA1972) 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 ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for TJP3 at approximately 130-140 kDa. The expected band size for TJP3 is at 101 kDa.
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IHC analysis of TJP3 using anti-TJP3 antibody (PA1972).
TJP3 was detected in a paraffin-embedded section of human fallopian tube tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-TJP3 Antibody (PA1972) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
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IF analysis of TJP3 using anti-TJP3 antibody (PA1972).
TJP3 was detected in an immunocytochemical section of CACO-2 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 5 μg/mL rabbit anti-TJP3 Antibody (PA1972) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:500 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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Flow Cytometry analysis of CACO-2 cells using anti-TJP3 antibody (PA1972).
Overlay histogram showing CACO-2 cells stained with PA1972 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-TJP3 Antibody (PA1972, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/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-Zonula occludens protein 3/TJP3 Antibody Picoband® (PA1972)
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