TJP1 antibody (Tight Junction Protein 1 (Zona Occludens 1)) (AA 1670-1720)

Details for Product anti-TJP1 Antibody No. ABIN675024, Supplier: Log in to see
Antigen
  • ZO-1
  • ZO1
  • zo1
  • tight junction protein 1
  • tight junction protein 1 (zona occludens 1)
  • zonula occludens 1
  • TJP1
  • Tjp1
  • ZO1
Alternatives
anti-Human TJP1 antibody for Immunofluorescence (fixed cells)
Epitope
AA 1670-1720
15
10
5
4
3
3
3
2
2
2
2
2
2
2
2
1
1
1
1
1
1
1
1
1
Reactivity
Human, Mouse (Murine), Pig (Porcine), Rat (Rattus)
90
32
32
19
11
5
4
4
3
2
1
1
1
1
Host
Rabbit
72
16
3
3
2
Clonality
Polyclonal
Conjugate
This TJP1 antibody is un-conjugated
3
2
2
2
1
1
1
1
1
1
1
1
1
1
1
1
1
Application
Immunofluorescence (Cultured Cells) (IF (cc)), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))
73
24
24
23
23
13
12
11
10
8
8
3
2
1
1
Options
Supplier
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Available images

'Independent Validation' Badge
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.
Primary Antibody
  • Antibody: human Tight Junction Protein 1 (Zona Occludens 1) (TJP1)
  • Catalog number: ABIN675024
  • Supplier: Log in to see
  • Supplier number: Log in to see
  • Lot number: 130913
Isotype Control Antibody
Secondary Antibody
  • Antibody: Biotinylated goat anti-rabbit/anti-mouse (Kit)
  • Catalog number: Log in to see
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  • Lot number: D05923BA
Controls
  • Tissues stained came from a human formalin-fixed paraffin embedded (FFPE) tissue microarray (12-003d):
  • Positive control (specimen known to contain the target protein): human testis, which is expected to express high levels of the antigen.
  • Negative Control (specimen known to not contain the target protein): Indeterminate; protein located on cytoplasmic membrane surface of intercellular tight junctions. The protein may be involved in signal transduction at cell-cell junctions. Expression is expected to be widespread. Thymus, expected to express much lower levels of the antigen as compared to testis, is shown to demonstrate a tissue with no detectable expression.
  • Primary antibody isotype control: Testis (specimen known to contain the target protein) treated with primary antibody isotype control instead of the primary antibody.
  • Secondary antibody only control: Testis (specimen known to contain the target protein) treated with secondary antibody only (no primary antibody).
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.
Validation Images
Immunohistochemistry validation image for anti-Tight Junction Protein 1 (Zona Occludens 1) (TJP1) (AA 1670-1720) antibody (ABIN675024) Figure 1: TJP1 immunostaining of human testis (brown). Counterstain in blue.
Immunohistochemistry validation image for anti-Tight Junction Protein 1 (Zona Occludens 1) (TJP1) (AA 1670-1720) antibody (ABIN675024) Figure 2: Isotype control immunostaining of human testis (brown). Counterstain in blue.
Immunohistochemistry validation image for anti-Tight Junction Protein 1 (Zona Occludens 1) (TJP1) (AA 1670-1720) antibody (ABIN675024) Figure 3: Secondary only control immunostaining of human testis (brown). Counterstain...
Immunohistochemistry validation image for anti-Tight Junction Protein 1 (Zona Occludens 1) (TJP1) (AA 1670-1720) antibody (ABIN675024) Figure 4: TJP1 immunostaining of human thymus (brown). Counterstain in blue.
Immunohistochemistry validation image for anti-Tight Junction Protein 1 (Zona Occludens 1) (TJP1) (AA 1670-1720) antibody (ABIN675024) Figure 4: Isotype control immunostaining of human thymus (brown). Counterstain in blue.
Immunohistochemistry validation image for anti-Tight Junction Protein 1 (Zona Occludens 1) (TJP1) (AA 1670-1720) antibody (ABIN675024) Figure 6: Secondary only immunostaining of human thymus (brown). Counterstain in blue.
Immunogen KLH conjugated synthetic peptide derived from human ZO-1
Isotype IgG
Purification Purified by Protein A.
Alternative Name Zo-1 (TJP1 Antibody Abstract)
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.

Subcellular location: Cytoplasm

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

Gene ID 7082
UniProt Q07157
Pathways Carbohydrate Homeostasis, Cell-Cell Junction Organization
Application Notes IHC-P 1:100-500
IF(IHC-P) 1:50-200
IF(ICC) 1:50-200
Restrictions For Research Use only
Format Liquid
Concentration 1 μg/μL
Buffer Aqueous buffered solution containing 1 % BSA, 50 % glycerol and 0.09 % sodium azide.
Preservative Sodium azide
Precaution of Use This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.
Storage -20 °C
Storage Comment Store at -20°C
Expiry Date 12 months
Supplier Images
Immunohistochemistry (IHC) image for anti-TJP1 antibody (Tight Junction Protein 1 (Zona Occludens 1)) (AA 1670-1720) (ABIN675024) Formalin-fixed and paraffin embedded rat brain labeled with Anti- ZO-1 Polyclonal Ant...
Immunohistochemistry (IHC) image for anti-TJP1 antibody (Tight Junction Protein 1 (Zona Occludens 1)) (AA 1670-1720) (ABIN675024) Image kindly submitted by Kamlesh Gupta as part of the free sample program. Caco-2 ce...
Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)) image for anti-TJP1 antibody (Tight Junction Protein 1 (Zona Occludens 1)) (AA 1670-1720) (ABIN675024) Image was kindly submitted by Sriram Srivatsa from Medical University of Vienna.Paraf...
Western Blotting (WB) image for anti-TJP1 antibody (Tight Junction Protein 1 (Zona Occludens 1)) (AA 1670-1720) (ABIN675024) Image kindly submitted by Kamlesh Gupta as part of the free sample program. Caco-2 ce...
Product cited in: 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).