TJP1 antibody (AA 1551-1702)

Catalog No. ABIN675024

Product Details anti-TJP1 Antibody

Target: TJP1 (Tight Junction Protein 1 (TJP1))

Binding Specificity: AA 1551-1702

Reactivity: Human, Mouse, Rat, Pig, Cow

Host: Rabbit

Clonality: Polyclonal

Conjugate: This TJP1 antibody is un-conjugated

Application: Western Blotting (WB), ELISA, Flow Cytometry (FACS), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunofluorescence (Cultured Cells) (IF (cc)), Immunohistochemistry (Frozen Sections) (IHC (fro))

Cross-Reactivity: Cow, Human, Mouse, Pig, Rat

Predicted Reactivity: Dog,Cow,Chicken,Rabbit,Guinea Pig

Purification: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human ZO-1

Isotype: IgG

Application Details

Application Notes: WB 1:300-5000
ELISA 1:500-1000
FCM 1:20-100
IHC-P 1:200-400
IHC-F 1:100-500
IF(IHC-P) 1:50-200
IF(IHC-F) 1:50-200
IF(ICC) 1:50-200

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #029577 (Immunohistochemistry)

by: Immunohistochemistry Core, NYU Langone

No.: #029577

Date: 01/18/2014

Lot Number: 130913

Method validated: Immunohistochemistry

Positive Control: Testis

Negative Control: See Controls section for negative controls

Notes: Signal was detected in positive control tissue, and not detected in a tissue expected to express very low levels of the target antigen.

Validation Images

Figure 1: TJP1 immunostaining of human testis (brown). Counterstain in blue.

Figure 2: Isotype control immunostaining of human testis (brown). Counterstain in blue.

Figure 3: Secondary only control immunostaining of human testis (brown). Counterstain in blue.

Figure 4: TJP1 immunostaining of human thymus (brown). Counterstain in blue.

Figure 4: Isotype control immunostaining of human thymus (brown). Counterstain in blue.

Figure 6: Secondary only immunostaining of human thymus (brown). Counterstain in blue.

Full Methods

Primary Antibody:

• Antibody: human Tight Junction Protein 1 (Zona Occludens 1) (TJP1)
• Catalog number: ABIN675024
• Lot number: 130913

Secondary Antibody:

• Antibody: Biotinylated goat anti-rabbit/anti-mouse (Kit)
• Lot number: D05923BA

Full Protocol:

• Immunohistochemistry was performed on a Ventana NexES automated platform, instrument manufacturer specific reagents are italicized.
• 1. Slides were preheated in convection oven at 60°C for 30 minutes
• 2. Deparaffinization procedure: - 3 changes of Xylene, 5 minutes each - 3 changes of 100% Ethanol, 3 minutes each - 3 changes of 95% Ethanol, 3 minutes each - Rinsed in distilled water, 3 changes
• 1. Heat retrieval procedure - Slides retrieved in 10.0 mM Citrate, pH6.0 in a 1000W microwave oven (~100°C) for 15 minutes. - Slides were allowed to cool (in citrate) for 30 minutes. - Slides were washed x 3 in Distilled water
• 1. NexES instrument procedure, iVIEW DAB paraffin protocol (*abridged*): - Slide chamber warmed to 37°C
• 1. Slides rinsed with *reaction buffer* x 3
• 1. *iVIEW Inhibitor (H2O2)* applied and incubated for 4 minutes
• 1. Slides rinsed with *reaction buffer*
• 1. Antibody Application - Primary antibody diluted 1:250 in PBS (100 microliters applied/slide) - Ventana Isotype control applied neat - Slides incubated overnight at room temperature (~12 hours ~25°C)
• 1. Slides rinsed with *reaction buffer* x3
• 1. *iVIEW Biotinylated IgG* applied and incubated for 8 minutes
• 1. Slides rinsed with *reaction buffer*
• 1. *iVIEW Streptavidin-Horseradish Peroxidase* applied and incubated for 8 minutes
• 1. Slides rinsed with *reaction buffer*
• 1. *iVIEW DAB/H2O2* applied and incubated for 8 minutes
• 1. Slides rinsed with *reaction buffer*
• 1. *iVIEW Copper* applied and incubated for 4 minutes
• 1. Slides rinsed with *reaction buffer*
• 1. Slides washed in Dawn Detergent/tap water
• 1. Counterstain Procedure - Hematoxylin (Leica 560 MX) 30 seconds - Slides washed in tap water, 1 minute - Decolorized (10% Acetic Acid in 70% ethanol), 1 minute - Slides washed in tap water, 1 minute - Bluing (Austin Clear Ammonia), 1 minute - Slides washed in tap water, 1 minute
• 1. Dehydration/coverslipping procedure: - 3 changes of 95% Ethanol, 3 minutes each - 3 changes of 100% Ethanol, 3 minutes each - 3 changes of Xylene, 5 minutes each - Mounted with Permount
• 1. Imaging: Leica SCN 400F Whole Slide Scanner with Digital Image Hub and Leica Slidepath software

Experimental Notes:

• Deviations from protocol/procedure supplied by manufacturer (attached).
• Step 1: Heated tissue 60°C for 30 minutes; manufacturer heats for 45 minutes.
• Step 2: No ethanol wash was performed during deparaffinization; manufacturer includes 1 wash of 80% ethanol for 3 minutes.
• Step 3.1: Slides were heated for 15 minutes; manufacturer provides a range of 15-20 minutes.
• Step 3.2: Slides were cooled for 30 minutes; manufacturer cools for 20 minutes.
• Step 4: Italicized reagents and incubation time are fixed instrument parameters.
• Step 5: Secondary species-specific serum block not used; manufacturer blocks with 5% normal goat serum for 2 hours.
• Step 8.1: Antibody diluted in PBS at 1:250; manufacture did not recommend diluent or dilution.
• Step 8.2.1: Primary antibody incubated at room temperature overnight; manufacturer incubates overnight 4°C with agitation.
• Tissue Interpretation (limited): - TJP1: Under the staining parameters described above, testis stained weakly (ducts) positive (Figure 1). Substantial signal detected in limited number of other tissues, including: breast, normal (NOS); pancreatic cancer (NOS), and stomach, normal (NOS). Most tissues showed low level of specific signal. Thymus did not have any detectable signal (Figure 4).
• I-NC (Isotype negative control): No signal detected
• B-NC (Blank negative control): No signal detected
• Signal Localization: - Signal to noise was adequate with cytoplasmic, nuclear subcellular localization observed. Rare inner-membrane and no distinct membrane signal observed.

Handling

Format: Liquid

Concentration: 1 μg/μL

Buffer: 0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.

Preservative: ProClin

Precaution of Use: This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.

Storage: 4 °C,-20 °C

Storage Comment: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Expiry Date: 12 months

References for anti-TJP1 antibody (ABIN675024)

Zhuang, Zhong, Bian, Fan, Chen, Liu, Liu: "Rhein ameliorates lipopolysaccharide-induced intestinal barrier injury via modulation of Nrf2 and MAPKs." in: Life sciences, Vol. 216, pp. 168-175, (2019) (PubMed).

Kulhankova, Kinney, Stach, Gourronc, Grumbach, Klingelhutz, Salgado-Pabón: "The Superantigen Toxic Shock Syndrome Toxin-1 Alters Human Aortic Endothelial Cell Function." in: Infection and immunity, (2018) (PubMed).

Zou, Lin, Li, Wu, He, Huang, Wang, Ye, Cheng, Ding, Zheng, Chi: "Huangqin-tang ameliorates dextran sodium sulphate-induced colitis by regulating intestinal epithelial cell homeostasis, inflammation and immune response." in: Scientific reports, Vol. 6, pp. 39299, (2018) (PubMed).

Zhao, Qin, Han, Wang, Zhang, Liu: "?-Conglycinin reduces the tight junction occludin and ZO-1 expression in IPEC-J2." in: International journal of molecular sciences, Vol. 15, Issue 2, pp. 1915-26, (2014) (PubMed).

Ruan, Liu, Zhou, Mi, Liu, Wu, Yao, Assaad, Deng, Hou, Wu, Yin: "Chlorogenic acid decreases intestinal permeability and increases expression of intestinal tight junction proteins in weaned rats challenged with LPS." in: PLoS ONE, Vol. 9, Issue 6, pp. e97815, (2014) (PubMed).

Zhao, Qin, Sun, Che, Bao, Zhang: "Effects of soybean agglutinin on intestinal barrier permeability and tight junction protein expression in weaned piglets." in: International journal of molecular sciences, Vol. 12, Issue 12, pp. 8502-12, (2012) (PubMed).

Gu, Xue, Wei, Zhang, Li: "Calcium-activated potassium channel activator down-regulated the expression of tight junction protein in brain tumor model in rats." in: Neuroscience letters, Vol. 493, Issue 3, pp. 140-4, (2011) (PubMed).

Target Details for TJP1

Target: TJP1 (Tight Junction Protein 1 (TJP1))

Alternative Name: ZO-1 (TJP1 Products)

Synonyms: ZO-1 antibody, ZO1 antibody, zo1 antibody, tight junction protein 1 antibody, tight junction protein ZO-1 antibody, TJP1 antibody, Tjp1 antibody, LOC396567 antibody

Background:

Synonyms: ZO-1, Tight junction protein ZO-1, Tight junction protein 1, Zona occludens protein 1, Zonula occludens protein 1, TJP1, ZO1

Background: The N-terminal may be involved in transducing a signal required for tight junction assembly, while the C-terminal may have specific properties of tight junctions. The alpha domain might be involved in stabilizing junctions. Plays a role in the regulation of cell migration by targeting CDC42BPB to the leading edge of migrating cells.

Gene ID: 7082

UniProt: Q07157

Pathways: Carbohydrate Homeostasis, Cell-Cell Junction Organization