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

Details for Product anti-TJP1 Antibody No. ABIN675024
Antigen
  • ZO-1
  • ZO1
  • zo1
  • tight junction protein 1
  • tight junction protein ZO-1
  • TJP1
  • Tjp1
  • LOC396567
Epitop
AA 1670-1720
15
10
4
4
3
3
3
3
2
2
2
2
2
2
1
1
1
1
1
1
1
Reaktivität
Human, Maus, Schwein, Ratte (Rattus)
116
56
42
20
13
7
6
6
5
3
3
2
2
2
2
2
2
1
1
1
1
Wirt
Kaninchen
96
18
4
2
1
Klonalität
Polyklonal
Konjugat
Dieser TJP1 Antikörper ist unkonjugiert
4
3
3
3
3
3
3
3
2
2
1
1
1
1
1
1
1
Applikation
Immunofluorescence (Cultured Cells) (IF (cc)), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))
86
35
32
22
20
13
12
12
12
8
8
6
2
1
1
Optionen
'Independent Validation' Siegel
Chargennummer 130913
Validierte Anwendung Immunohistochemistry
Positivkontrolle Testis
Negativkontrolle See Controls section for negative controls
Bewertung Signal was detected in positive control tissue, and not detected in a tissue expected to express very low levels of the target antigen.
Primärantikörper
  • Antibody: human Tight Junction Protein 1 (Zona Occludens 1) (TJP1)
  • Catalog number: ABIN675024
  • Lot number: 130913
Isotypkontrolle
  • Antibody: Rabbit IgG isotype control
  • Lot number: C11487
Sekundärantikörper
  • Antibody: Biotinylated goat anti-rabbit/anti-mouse (Kit)
  • Lot number: D05923BA
Kontrollen
  • 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).
Protokoll
  • 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
Anmerkungen
  • 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.
Validierungsbilder
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
Isotyp IgG
Reinigung Purified by Protein A.
Plasmids, Primers & others Plasmide, Primers & weitere TJP1 products on genomics-online (e.g. as negative or positive controls)
Antigen
Andere Bezeichnung Zo-1 (TJP1 Antibody Abstract)
Hintergrund

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

Gen-ID 7082
UniProt Q07157
Pathways Carbohydrate Homeostasis, Cell-Cell Junction Organization
Applikations-hinweise IHC-P 1:100-500
IF(IHC-P) 1:50-200
IF(ICC) 1:50-200
Beschränkungen Nur für Forschungszwecke einsetzbar
Format Liquid
Konzentration 1 μg/μL
Buffer Aqueous buffered solution containing 1 % BSA, 50 % glycerol and 0.09 % sodium azide.
Konservierungs-mittel Sodium azide
Vorsichtsmaßnahmen This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.
Lagerung -20 °C
Informationen zur Lagerung Store at -20°C
Haltbarkeit 12 months
Bilder des Herstellers
Immunohistochemistry (IHC) image for anti-Tight Junction Protein 1 (Zona Occludens 1) (TJP1) (AA 1670-1720) antibody (ABIN675024) Formalin-fixed and paraffin embedded rat brain labeled with Anti- ZO-1 Polyclonal Ant...
Immunohistochemistry (IHC) image for anti-Tight Junction Protein 1 (Zona Occludens 1) (TJP1) (AA 1670-1720) antibody (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-Tight Junction Protein 1 (Zona Occludens 1) (TJP1) (AA 1670-1720) antibody (ABIN675024) Image was kindly submitted by Sriram Srivatsa from Medical University of Vienna.Paraf...
Western Blotting (WB) image for anti-Tight Junction Protein 1 (Zona Occludens 1) (TJP1) (AA 1670-1720) antibody (ABIN675024) Image kindly submitted by Kamlesh Gupta as part of the free sample program. Caco-2 ce...
Produkt verwendet in: 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). Von den Autoren verwendete Methode: Flow Cytometry (FACS) (Probematerial (Species): Human).

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). (Probematerial (Species): Pig (Porcine)).

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). Von den Autoren verwendete Methode: Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)) (Probematerial (Species): Rat (Rattus)).

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). Von den Autoren verwendete Methode: Western Blotting (WB), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)) (Probematerial (Species): Pig (Porcine)). Weitere Details: Sample: Mid-jejunum fragments

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). Von den Autoren verwendete Methode: Western Blotting (WB) (Probematerial (Species): Rat (Rattus)). Weitere Details: Sample: brain

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