- PA5-28858 - Invitrogen Antibodies TJP1 antibody

Submitted references Curcumin reduces blood-nerve barrier abnormalities and cytotoxicity to endothelial cells and pericytes induced by cisplatin.

Kobutree P, Tothonglor A, Roumwong A, Jindatip D, Agthong S
Folia morphologica 2022 Jul 12;

miR-4456/CCL3/CCR5 Pathway in the Pathogenesis of Tight Junction Impairment in Chronic Obstructive Pulmonary Disease.

Yu W, Ye T, Ding J, Huang Y, Peng Y, Xia Q, Cuntai Z
Frontiers in pharmacology 2021;12:551839

Hyaluronan 35 kDa enhances epithelial barrier function and protects against the development of murine necrotizing enterocolitis.

Gunasekaran A, Eckert J, Burge K, Zheng W, Yu Z, Kessler S, de la Motte C, Chaaban H
Pediatric research 2020 Jun;87(7):1177-1184

Protective effect of Saccharomyces boulardii on intestinal mucosal barrier of dextran sodium sulfate-induced colitis in mice.

Dong JP, Zheng Y, Wu T, He Q, Teng GG, Wang HH
Chinese medical journal 2019 Aug 20;132(16):1951-1958

Anesthesia and Surgery Impair Blood-Brain Barrier and Cognitive Function in Mice.

Yang S, Gu C, Mandeville ET, Dong Y, Esposito E, Zhang Y, Yang G, Shen Y, Fu X, Lo EH, Xie Z
Frontiers in immunology 2017;8:902

RFX2 Is a Major Transcriptional Regulator of Spermiogenesis.

Kistler WS, Baas D, Lemeille S, Paschaki M, Seguin-Estevez Q, Barras E, Ma W, Duteyrat JL, Morlé L, Durand B, Reith W
PLoS genetics 2015 Jul;11(7):e1005368

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of ZO-1 using Various whole cell extracts (30 µg). Samples were loaded onto a 5% SDS-PAGE gel and probed with a ZO-1 polyclonal antibody (Product # PA5-28858) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of ZO-1 using Various whole cell extracts (30 µg). Samples were loaded onto a 5% SDS-PAGE gel and probed with a ZO-1 polyclonal antibody (Product # PA5-28858) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of ZO-1 was performed by separating 30 µg of various whole cell extracts by 5% SDS-PAGE. Proteins were transferred to a membrane and probed with a ZO-1 Polyclonal Antibody (Product # PA5-28858) at a dilution of 1:500 and a HRP-conjugated anti-rabbit IgG secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using ZO-1 Polyclonal Antibody (Product # PA5-28858). Mouse tissue extract (50 µg) was separated by 5% SDS-PAGE, and the membrane was blotted with ZO-1 Polyclonal Antibody (Product # PA5-28858) diluted at 1:500. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of ZO-1 was performed by separating 30 µg of various whole cell extracts by 5% SDS-PAGE. Proteins were transferred to a membrane and probed with a ZO-1 Polyclonal Antibody (Product # PA5-28858) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of ZO-1 was performed using 90% confluent log phase Caco-2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with ZO-1 Polyclonal Antibody (Product # PA5-28858) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing Junctional localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of ZO-1 was achieved by transfecting Caco-2 cells with ZO-1 specific siRNA (Silencer® select Product # s14155, s14156). Immunofluorescence analysis was performed on untransfected Caco-2 cells (panel a-d), transfected with non-specific scrambled siRNA (panels e-h) and transfected with ZO-1 specific siRNA (panel i-l). Cells were fixed, permeabilized, and labelled with ZO-1 Polyclonal Antibody (Product # PA5-28858, 1:100 dilution) followed by Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution). Nuclei (blue) were stained using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962), and Rhodamine Phalloidin (Product # R415, 1:300 dilution) was used for cytoskeletal F-actin (Red) staining. Reduction in signal of specific signal was observed upon siRNA mediated knockdown (panel i-l) confirming specificity of the antibody to ZO-1 (Green). The Images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry (Paraffin) analysis of ZO-1 was performed in paraffin-embedded human breast carcinoma tissue using ZO-1 Polyclonal Antibody (Product # PA5-28858) at a dilution of 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry (Paraffin) analysis of ZO-1 was performed in paraffin-embedded mouse cervix tissue using ZO-1 Polyclonal Antibody (Product # PA5-28858) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 2 CCL3 promotes epithelial tight junction injury by binding with CCR5 (A) Tight junction proteins ZO-1 (red), occludin (green), and merged (blue + orange) were stained in 16HBE cells (left panel) and A549 cells (right panel) by immunofluorescence. Presented as means +- standard error. n = 3, * p < 0.05, vs. the control group. Data are representative of three independent experiments. Scale bar = 10 um (B) Transepithelial electrical resistance (TER) after CCL3 treatment (10 mg/ml), as a cell function test 7 days after plating the airway epithelial cells on coated permeable filters. Presented as means +- standard error. n = 3, * p < 0.05, vs. the control group (C) Western blot analysis to detect protein expression of ZO-1, claudin and occludin in 16HBE cells and A549 cells using different concentrations of CCL3. n = 3, * p < 0.05, vs. the control group, ** p < 0.01, vs. the control group (CCL3: 0 ng/ml). The expression levels were normalized to the expression of beta-actin (left panel). n = 3 (D) Real-time PCR assays of ZO-1 (upper) and occludin (lower) mRNA expression in 16HBE cells and A549 cells using different concentrations of CCL3. n = 3, * p < 0.05, vs. the Ctrl group, ** p < 0.01, vs. the control group. Protein expression was normalized to GAPDH (E) Tight junction proteins ZO-1 (red), occludin (green) and merged (blue and orange) were stained by immunofluorescence in the four groups: 1) Ctrl (PBS:10 mg ml -1 ), 2) CSE (10 mg ml -1 ), 3) CCL3 (CCL3 10 mg ml -1 ), a

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 3 Identification of miR-4456 as an upstream signal of CCL3 induced-tight junction injury (A) Real-time PCR screen in 16-HBE cells (left panel) and A549 cells (right panel) for miRNAs that could target CCL3 mRNA. CCL3 mRNA expression levels were normalized to GAPDH expression. n = 3, * p < 0.05, vs. the control group. The expression levels were normalized to the expression of GAPDH (B) Baseline expression of miR-4456 according to COPD grade. * p < 0.05 vs. the non-smoker group (( n = 48. non-smokers:8, GOLD 1:9, GOLD2: 9, GOLD3:10, GOLD4:12) (C) Western blot analysis of ZO-1, occludin, CCL3, and CCR5 protein expression in the four groups: 1) Control (PBS 10 mg ml -1 ), 2) CSE (10 mg ml -1 ), 3) miR-4456, and 4) CSE (10 mg ml -1 )+miR-4456 mimics in 16HBE cells and A549 cells. n = 3, ** p < 0.01, vs. the control group, * p < 0.05, vs. the control group, # p < 0.05, vs. the CSE group. Protein expression was normalized to beta-actin (D) Correlation analysis of miR-4456 and CCL3 mRNA levels by Pearson correlation coefficient from a total of 22 COPD peripheral blood samples ( n = 22) (E) Tight junction proteins ZO-1 (red), occludin (green) and merged (blue + orange) were stained by immunofluorescence in the four groups: 1) Control (PBS 10 mg ml -1 ), 2) CSE (10 mg ml -1 ), 3) CSE (10 mg ml -1 )+miR4456, and 4) CSE (10 mg ml -1 )+miR4456 mimics + CCL3 (10 mg ml -1 ) in 16HBE cells (left panel) and A549 cells (right panel). Presented as means +- standard error. n = 3, * p < 0.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 5 In vivo mouse studies confirm the role of CCL3 and miR-4456 in COPD (A) H&E staining confirms the macroscopic appearance of pulmonary tissue in the four groups: 1) Control (Ctrl), 2) CSE, 3) CCL3, and 4) CSE + DAPTA. Quantification of the alveolar space is in the lower panels. n = 8,* p < 0.05, vs. the control group, # p < 0.05, vs. the CSE group (B) Airway resistance, Elasticity, Compliance, and total BALF proteins were detected in the four groups same as Figure 5A n = 8,* p < 0.05, vs. the control group, # p < 0.05, vs. the CSE group (C) Inflammatory and oxidative stress markers in the four groups same as Figure 5A n = 8,* p < 0.05, vs. the control group, # p < 0.05, vs. the CSE group (D) Tight junction proteins ZO-1 merge (red and blue), and occludin merge (green and blue) were stained using immunofluorescence in the four groups same as Figure 5A. Data are representative of three independent experiments. Scale bar = 10 um n = 8, * p < 0.05, vs. the control group, # p < 0.05, vs. the CSE group (E) Real-time PCR assays were performed in the blood and lung to detect ZO-1 and occludin mRNA before and after CSE treatment in the four groups same as Figure 5A. Data are represented as the mean +- standard deviation. n = 8, * p < 0.05, vs. the control group. # p < 0.05, vs. the CSE group. The expression levels were normalized to the expression of GAPDH, n = 3 (F) Western blot for ZO-1 and occludin protein expression in the four groups same as Figure 5A n = 8, * p < 0.05, v

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2 Immunohistochemical staining of ZO-1 from normal control group (A), colitis model group (B), Sb treatment group (C), mesalazine treatment group (D), and combination treatment group (E) (Original magnification x400). The mean ZO-1 score from group A to group E (F). * P < 0.01, + P < 0.05. Sb: Saccharomyces boulardii ; ZO-1: Zona occludens.

PA5-28858

Antibody data