35-2500

antibody from Invitrogen Antibodies

Targeting: CLDN5 AWAL, BEC1, CPETRL1, TMVCF

Provider product page for 35-2500

Western blot

ELISA

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [227]
Comments [0]
Validations
Western blot [2]
Immunohistochemistry [1]
Other assay [7]

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Product number: 35-2500 - Provider product page
Provider: Invitrogen Antibodies
Product name: Claudin 5 Monoclonal Antibody (4C3C2)
Antibody type: Monoclonal
Antigen: Synthetic peptide
Reactivity: Human, Mouse, Rat
Host: Mouse
Isotype: IgG
Antibody clone number: 4C3C2
Vial size: 100 µg
Concentration: 0.5 mg/mL
Storage: -20°C

CLDN5 protein structure - 35-2500 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of Claudin-5 was performed by loading 20 µg of Mouse Placenta (lane1) and Mouse Small Intestine (lane2) lysates using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and iBlot® Dry Blotting System (IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5 % skim milk for 1 hour at room temperature. Claudin-5 was detected at ~23 kDa using Claudin-5 Mouse Monoclonal Antibody (Product # 35-2500) at 0.5-1 µg/mL in 2.5 % skim milk at 4°C overnight on a rocking platform. Goat Anti-Mouse IgG - HRP Secondary Antibody (Product # 62-6520) at 1:4000 dilution was used and chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-Claudin 5 Monoclonal Antibody (Product # 35-2500) and an 18kDa band corresponding to Claudin 5 was observed to be downregulated upon treatment with VEGF and TGF beta. In addition, expression of Claudin 5 was observed only in HUVEC as compared to THP-1 and MOLT-4 which are reported to be negative. Membrane enriched extracts (30 µg lysate) of HUVEC (Lane 1), HUVEC treated with VEGF (5ng/ml for 48 hours) (Lane 2), HUVEC treated with TGF beta (30ng/ml for 48 hours) (Lane 3), THP-1 (Lane 4) and MOLT-4 (Lane 5) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005)..

Supportive validation

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

Supportive validation

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

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 3 Protein expression (Western blot) of claudin-1, claudin-3, claudin-4, claudin-5, claudin-7, and claudin-8 after treatment with bacterial strains (Ecf, ETEC, and Ecf + ETEC). (a) Protein expression relative to respective controls [means +- SEM]. Samples were taken after 8 h. No differences were observed between treatment groups. N = 3 independent experiments per bar. (b) Exemplarily, data of one Western blot is shown. 1 = control (8 h), 2 = Ecf (8 h), 3 = ETEC (8 h), 4 = Ecf (8 h) + ETEC (6 h), and 5 = Ecf (10 h) + ETEC (8 h). Sample 4 was included as a control to rule out effects of longer total bacterial incubation (preincubation) and was not included in the statistical analysis shown in (a).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 1 Expression of TJPs in different leukocyte subsets. Immunofluorescence staining of PBLs from healthy controls with antibodies against claudin 1 or claudin 5 (green) and CD20 (A,B), CD14 (C,D), CD3 (E,F), CD4 (G,H), CD8 (I,J), ZO-1 (K,L) and ZO-2 (M,N) (all red) and a nuclear dye (TO-PRO 3-blue). Scale bar: 5 mum. Representative results from three independent experiments are shown.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 6 Claudin-5 and claudin-4 are expressed in endothelial cells in the developing brain a - d . Cld-5 immunoreactivity in the developing and adult brain. Parenchymal vessels are labeled ( green ) in E19 ( arrows a ) or adult ( arrows b , d ) animals. In c , the anti-Cld-5 polyclonal antibody was used to show the colocalization of the TJ protein ( red ) with the endothelial marker RECA-1 ( green ) in a P2 animal. In all CPs immunodetection revealed a signal associated with the largest vessels penetrating the choroidal stroma ( arrowheads b , d ), but not with the choroidal epithelium, labeled in red by anti-Cld-1 Ab ( d ). Note that all cisternal vessels such as the vein of Galen are also labeled by anti-Cld-5 Ab. DAPI was used for nuclei staining. e Representative Western blot of Cld-4. Brain cortex ( lane 1 20 mug) and MV ( lanes 2 and 3 40 mug) were isolated from adult animals. Lane 4 was loaded with MDCK (10 mug) as positive control. A low, but specific Cld-4 signal was detected in MV ( upper panel lane 3 band at 22 kDa), but not in cortex or in MV when the primary antibody was omitted ( lane 2 ). Actin used as a loading indicator is shown in the lower panel (42 kDa band). Due to the high expression of Cld-4 in MDCK cells, lane 4 was treated separately from the other lanes for HRP detection. f - h Cld-4 immunoreactivity ( red ) in the developing and adult brain. Double immunolabeling with anti-RECA-1Ab ( green ) allowed visualizing the parenchymal vessels that were posit

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 8 Representative immunofluorescence images of dhBMEC monolayers fixed and stained after 40 h at 0, 4, and 12 dyne cm -2 . a , e , i CLDN-5. b , f , j OCLN. c , g , k ZO-1. d , h , l F-actin. Note that CLDN-5/OCLN and ZO-1/f-actin were obtained for different monolayers