Invitrogen

Claudin 18 Recombinant Rabbit Monoclonal Antibody (34H14L15)

Product Image Gallery

Claudin 18 Antibody in Western Blot (WB) — Western blot analysis of Claudin-18 was performed by loading 30 µg of rat Heart, A431 and A549 lysates using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (Product # LC5800), and iBlot® Dry Blotting System (Product # IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5% skim milk for 1 hour at room temperature. Claudin-18 was detected at ~27 kDa using Claudin-18 Recombinant Rabbit Monoclonal Antibody (Product # 700178) at a 1:1000 dilution in 2.5% skim milk at 4°C overnight on a rocking platform. Detection was performed using an HRP-conjugated Goat anti-Rabbit secondary antibody (Product # G-21234) at a 1:5000 dilution and chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Claudin 18 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of Claudin-18 was performed on paraffin-embedded human normal stomach and squamous lung carcinoma tissue sections. To expose target proteins, endogenous peroxide blocking followed by heat induced epitope retrieval was performed using Tris-EDTA (pH 9. 0) buffer for 15 minutes. Following antigen retrieval, tissues were blocked in 0. 2% BSA with 0. 1% cold fish skin gelatin in 1X PBS for 1 hour in a humidified chamber. The tissues were then probed with a Claudin-18 Recombinant Rabbit Monoclonal Antibody (Product # 700178) at a dilution of 2 µg/mL for 3 hours at room temperature in a humidified chamber. Detection was performed using an HRP-conjugated Goat anti-Rabbit secondary antibody (Product # G-21234), followed by colorimetric detection using DAB. Images were taken at 40x (normal stomach) or 20x (lung carcinoma) magnification. Note membrane staining of normal stomach and cytoplasmic staining of lung carcinoma.

Claudin 18 Antibody in Immunofluorescence (IF) — Immunofluorescent analysis of Claudin-18 was done on 90% confluent log phase Caco-2 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with Claudin-18 Recombinant Rabbit Monoclonal Antibody (Product # 700178) at a dilution of 1:500 in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor® 488 Goat anti-Rabbit IgG Secondary Antibody (Product # A-11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c is a merged image showing cell junction localization and panel d is a no primary antibody control. The images were captured using a Nikon microscope at 20X magnification.

Claudin 18 Antibody — Figure 7 Ductal deletion of Hnf1b leads to PanIN by 2 months. ( A ) H and E staining of mutant pancreata showing epithelial structures composed of columnar cells with abundant supranuclear cytoplasm and basally located nuclei. ( B ) Alcian blue staining. Columnar mutant epithelial cells showed blue-stained supranuclear mucin. ( C ) PanIN marker claudin 18 (brown) immunohistochemistry. ( D ) Sox9 (red) and GFP (green) immunostaining. Sox9+ ADM structures did not derive from Hnf1b-targeted GFP+ cells. ( E ) Quantification of the relative surface of acini, adipocytes, fibrosis/infiltrates, ADM, and PanINs in mutants (n = 6). Scale bars : 100 mum. Control, n >= 3; mutant, n >= 3 for histology and immunohistochemistry. DAPI, 4',6-diamidino-2-phenylindole.

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

2-3 µg/mL

View 4 publications 4 publications

Immunohistochemistry (IHC)

2-4 µg/mL

View 8 publications 8 publications

Immunohistochemistry (Paraffin) (IHC (P))

2-4 µg/mL

View 2 publications 2 publications

Immunocytochemistry (ICC)

1:100-1:1000

View 1 publication 1 publication

Immunofluorescence (IF)

1:100-1:1000

View 3 publications 3 publications

Immunoprecipitation (IP)

View 1 publication 1 publication

Product Specifications

Species Reactivity

Human, Mouse, Rat

Published species

Human, Mouse, Rat

Host/Isotype

Rabbit / IgG

Class

Recombinant Monoclonal

Type

Antibody

Clone

34H14L15

Immunogen

A peptide corresponding to amino acids 224-237 of P56856.

Conjugate

Unconjugated

Form

Liquid

Concentration

0.5 mg/mL

Purification

Protein A

Storage buffer

PBS

Contains

0.09% sodium azide

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RRID

AB_2532290

Product Specific Information

This antibody is predicted to react with chimpanzee, equine, rat and Rhesus monkey based on sequence homology.

Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.The immunogenic sequence for this antibody is GFKASTGFGSNTKNH, and the antibody is predicted to react with chimpanzee, equine, rat, and Rhesus monkey based on sequence identity and similarity.

Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.

Target Information

Claudin-18 is a tight junction protein expressed as lung-and stomach-specific isoforms which are generated by alternative splicing of the claudin-18 gene. The lung-specific form is a downstream target gene regulated by the T/EBP/NKX2.1 transcription factor. A splice variant lacking the C-terminal cytoplasmic domain also exists in mouse, but has not been confirmed in human. Human claudin-18 demonstrates 88% amino acid sequence identity to the mouse protein. Immunohistochemical studies have demonstrated complete membrane localization of claudin-18 in lung and stomach epithelial cells, while electron microscopy has shown that it is concentrated in the cell-cell borders of these cells. These features suggest a potentially important role for claudin-18 in the structure and function of tight junctions in the lung and stomach. Claudin-18 expression has also been reported in the inner ear.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: claudin 18-like; Claudin-18; surfactant associated 5; surfactant associated protein J; surfactant, pulmonary associated protein J

Gene Aliases: CLDN18; SFTA5; SFTPJ; UNQ778/PRO1572

UniProt ID: (Human) P56856, (Mouse) P56857

Entrez Gene ID: (Human) 51208, (Mouse) 56492, (Rat) 315953

Molecular Function: cell junction protein tight junction