Anti-ZO1 tight junction protein/TJP1 Antibody Picoband™

Product Name

Anti-ZO1 tight junction protein/TJP1 Antibody Picoband™

View all Tight Junction Protein 1 Antibodies

SKU/Catalog Number

PB9234

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-ZO1 tight junction protein/TJP1 Antibody Picoband™ catalog # PB9234. Tested in Flow Cytometry, IF, IHC, 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-ZO1 tight junction protein/TJP1 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9234)

Host

Rabbit

Contents

Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human TJP1 recombinant protein (Position: H1178-F1527). Human TJP1 shares 82% amino acid (aa) sequence identity with mouse TJP1.

*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

PB9234 is reactive to TJP1 in Human, Mouse, Rat

Applications

PB9234 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee

Observed Molecular Weight

220 kDa

Calculated molecular weight

195.459kDa

Background of Tight Junction Protein 1

Tight junction protein ZO-1 is a protein that in humans is encoded by the TJP1 gene. It is mapped to 15q13.1. This gene encodes a protein located on a cytoplasmic membrane surface of intercellular tight junctions. The encoded protein may be involved in signal transduction at cell–cell junctions. It has been found that injected CagA associates with the epithelial tight-junction scaffolding protein TJP1 and the transmembrane protein junctional adhesion molecule, causing an ectopic assembly of tight junction components at sites of bacterial attachment, and altering the composition and function of the apical-junctional complex.

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, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, By Heat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry, 1-3μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of TJP1 using anti-TJP1 antibody (PB9234).
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 30 ug of sample under reducing conditions.
Lane 1: human PC-3 whole cell lysates,
Lane 2: human CACO-2 whole cell lysates,
Lane 3: human COLO320 whole cell lysates,
Lane 4: rat testis tissue lysates,
Lane 5: mouse testis tissue 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-TJP1 antigen affinity purified polyclonal antibody (Catalog # PB9234) 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 TJP1 at approximately 220 kDa. The expected band size for TJP1 is at 185 kDa.

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Figure 2. IHC analysis of TJP1 using anti-TJP1 antibody (PB9234).
TJP1 was detected in paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-TJP1 Antibody (PB9234) 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. IF analysis of TJP1 using anti-TJP1 antibody (PB9234).
TJP1 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 2μg/mL rabbit anti-TJP1 Antibody (PB9234) 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 4. Flow Cytometry analysis of K562 cells using anti-TJP1 antibody (PB9234).
Overlay histogram showing K562 cells stained with PB9234 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TJP1 Antibody (PB9234,1μg/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 TJP1 (Source: Uniprot.org, NCBI)

Alternative Names

DKFZp686M05161; MGC133289; tight junction protein 1 (zona occludens 1); Tight junction protein 1; tight junction protein ZO-1; TJP1; ZO1; ZO-1; zona occludens 1; Zona occludens protein 1; zonula occludens 1 protein; Zonula occludens protein 1 TJP1 ZO-1 tight junction protein 1 tight junction protein ZO-1|zona occludens 1|zonula occludens 1 protein

For more info on TJP1, check out the TJP1 Infographic

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In this infographic, you will see the following information for TJP1: 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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