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Purified anti-CGN Antibody

Pricing & Availability
Clone
W21116A (See other available formats)
Regulatory Status
RUO
Other Names
Cingulin, KIAA1319
Isotype
Rat IgG2a, κ
Ave. Rating
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Product Citations
publications
a. W21116A_PURE_CGN_WB_100923
Whole cell extracts (15 µg total protein per lane) from the indicated cell lines were resolved on a 4-12% Bis-Tris gel, transferred to a PVDF membrane, and probed with 1.0 µg/mL of Purified anti-CGN (clone W21116A) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-rat IgG (Cat. No. 405405) at a 1:5000 dilution. Direct-Blot™ HRP anti-β-actin (Cat. No. 664804) was used as a loading control at a 1:25000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • a. W21116A_PURE_CGN_WB_100923
    Whole cell extracts (15 µg total protein per lane) from the indicated cell lines were resolved on a 4-12% Bis-Tris gel, transferred to a PVDF membrane, and probed with 1.0 µg/mL of Purified anti-CGN (clone W21116A) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-rat IgG (Cat. No. 405405) at a 1:5000 dilution. Direct-Blot™ HRP anti-β-actin (Cat. No. 664804) was used as a loading control at a 1:25000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • b. W21116A_PURE_CGN_ICC_100923
    C2BBe1 cells were fixed with Fixation Buffer (Cat. No. 420801) for 10 minutes, permeabilized with 0.5% Triton X-100, and blocked with 5% FBS for 1 hour at room temperature. Cells were then intracellularly stained with 2.0 µg/mL of Purified anti-CGN (clone W21116A), followed by incubation with 2.5 µg/mL of Alexa Fluor® 488 Goat anti-rat IgG (Cat. No. 405418) (green) for 1 hour at room temperature (panel A). Nuclei were counterstained with DAPI (Cat. No. 422801) (blue), and F-actin was counterstained with Flash Phalloidin™ Red 594 (Cat. No. 424203) (panel B). The image was captured with a 40X objective. Scale bar: 25 µm
  • c. W21116A_PURE_CGN_KO-KD_100923
    HeLa cells (mock) (panel A and B) and HeLa knockdown cells (panel C and D) were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with ice-cold methanol, and blocked with 5% FBS for 1 hour at room temperature. Cells were then intracellularly stained with 5.0 µg/mL of Purified anti-CGN (clone W21116A), followed by incubation with 2.5 µg/mL of Alexa Fluor® 594 Goat anti-rat IgG (Cat. No. 405422) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801) (panels B and D), and the image was captured with a 40X objective. Scale bar: 25 µm
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624951 25 µg $230
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624952 100 µg $455
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Description

CGN, found in the cytoplasmic plaque of tight junctions, is situated on the cytoplasmic side of tight junctions in polarized epithelia and certain endothelia. It is made up of a homodimer structure, with each subunit having a globular "head" section, a coiled-coil "rod" section, and a tiny globular "tail" section. Furthermore, CGN binds to JAM, ZO proteins, AF-6, and myosin, indicating its potential role in transmitting the mechanical energy produced by the contraction of the actomyosin cytoskeleton to the proteins of the cytoplasmic plaque and the membrane regions of TJs. Its main function is likely in the formation and control of the tight junction paracellular permeability barrier. Cingulin has additionally been discovered to engage with microtubules via the N-terminal head area, and these connections were governed by the process of phosphorylation by the adenosine monophosphate-activated protein kinase (AMPK). Moreover, the production of CGN in initial mouse embryos is specific to certain tissues, and it takes place in the blastocyst (increased in trophectoderm and decreased in inner-cells).

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Partial Recombinant human CGN protein
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is  0.125 - 1.0 µg/mL. For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/mL is recommended. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

The clone does not cross-react with mouse.

For immunocytochemistry (ICC), the suggested fixation/permeabilization method is either 4% paraformaldehyde/0.5% Triton X-100 or paraformaldehyde followed by ice-cold methanol, or fixed with ice-cold methanol only.

This product is not recommended for use in immunohistochemistry.

Additional Product Notes

This antibody has been tested in knockout/knockdown models for Western Blotting and Immunocytochemistry.

RRID
AB_3097248 (BioLegend Cat. No. 624951)
AB_3097248 (BioLegend Cat. No. 624952)

Antigen Details

Structure
CGN protein is a 1203 amino acid protein with a predicted molecular weight of 137 kD
Distribution

Pancreas, kidney, liver and lung, but not in skeletal muscle, placenta, brain or heart

Function
Formation and regulation of the tight junction
Interaction
TJP1/ZO1, SPEF1
Cell Type
Endothelial cells, Epithelial cells
Biology Area
Cell Biology, Cell Motility/Cytoskeleton/Structure
Antigen References
  1. Cordenonsi M, et al. 1999. J Cell Biol.  147:1569–82.
  2. Bazzoni G, et al. 2000. J Biol Chem. 275:20520-6.
  3. Yano T, et al. 2013. J Cell Biol. 203: 605–14.
  4. Javed Q, et al. 1993. Development. 117:1145-51.
  5. Fleming TP, et al. 1993. Development. 117:1135-44.
Gene ID
57530 View all products for this Gene ID
UniProt
View information about CGN on UniProt.org

Related FAQs

There are no FAQs for this product.
Go To Top Version: 1    Revision Date: 10/09/2023

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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