mouse clc 2  (Alomone Labs)


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    Name:
    Anti CLC 2 CLCN2 Antibody
    Description:
    Anti CLC 2 CLCN2 Antibody ACL 002 is a highly specific antibody directed against an epitope of the rat protein The antibody can be used in western blot immunohistochemistry and immunocytochemistry applications It has been designed to recognize CLC 2 channel from rat mouse and human samples
    Catalog Number:
    ACL-002
    Price:
    495.0
    Category:
    Primary Antibody
    Applications:
    Immunocytochemistry, Immunofluorescence, Immunohistochemistry, Western Blot
    Purity:
    Affinity purified on immobilized antigen.
    Immunogen:
    Synthetic peptide
    Size:
    50 mcl
    Antibody Type:
    Polyclonal Primary Antibodies
    Format:
    Lyophilized Powder
    Host:
    Rabbit
    Isotype:
    Rabbit IgG
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    Structured Review

    Alomone Labs mouse clc 2
    Anti CLC 2 CLCN2 Antibody
    Anti CLC 2 CLCN2 Antibody ACL 002 is a highly specific antibody directed against an epitope of the rat protein The antibody can be used in western blot immunohistochemistry and immunocytochemistry applications It has been designed to recognize CLC 2 channel from rat mouse and human samples
    https://www.bioz.com/result/mouse clc 2/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse clc 2 - by Bioz Stars, 2021-09
    93/100 stars

    Images

    1) Product Images from "Clcn2 encodes the hyperpolarization-activated chloride channel in the ducts of mouse salivary glands"

    Article Title: Clcn2 encodes the hyperpolarization-activated chloride channel in the ducts of mouse salivary glands

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    doi: 10.1152/ajpgi.90384.2008

    Functional expression of the ClC-2 channel in submandibular granular duct cells. A : differential interference contrast images of single granular duct and acinar cells freshly isolated from male mouse submandibular glands. Scale bar = 10 μm
    Figure Legend Snippet: Functional expression of the ClC-2 channel in submandibular granular duct cells. A : differential interference contrast images of single granular duct and acinar cells freshly isolated from male mouse submandibular glands. Scale bar = 10 μm

    Techniques Used: Functional Assay, Expressing, Isolation

    ClC-2 distribution in mouse submandibular and parotid glands. A : immunoperoxidase labeling for ClC-2 in the submandibular gland of a Clcn2 +/+ male mouse. AC, acinar cells; GD, granular duct; SD, striated duct; ED, excretory duct. Inset,
    Figure Legend Snippet: ClC-2 distribution in mouse submandibular and parotid glands. A : immunoperoxidase labeling for ClC-2 in the submandibular gland of a Clcn2 +/+ male mouse. AC, acinar cells; GD, granular duct; SD, striated duct; ED, excretory duct. Inset,

    Techniques Used: Labeling

    2) Product Images from "Epithelial Biology and Secretion: Knockout of ClC-2 reveals critical functions of adherens junctions in colonic homeostasis and tumorigenicity"

    Article Title: Epithelial Biology and Secretion: Knockout of ClC-2 reveals critical functions of adherens junctions in colonic homeostasis and tumorigenicity

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    doi: 10.1152/ajpgi.00087.2018

    Chloride channel protein-2 (ClC-2) knockout (KO) mice have shown disrupted adherens junctions (AJs) with nuclear localization of β-catenin in colon compared with wild-type (WT) mice. Sections of colonic tissue of ClC-2 WT and KO mice were immunolabeled for E-cadherin and β-catenin (red), and the nucleus (blue). A : ClC-2 KO colonic crypts showed dramatic disruptions of E-cadherin in upper crypts and elevated cytosolic and nuclear distribution of β-catenin. B : proximity ligation assay (PLA) showing the interaction (red dots) between E-cadherin and β-catenin in ClC-2 KO mouse colonic crypts was significantly reduced compared with ClC-2 WT. C : nuclear β-catenin was measured using ImageJ and showed significant elevations in ClC-2 KO colon ( n = 6). D : mRNA from the colonic tissues of WT and KO mice were studied for expression of T cell factor-1/lymphoid-enhancing factor-1 (TCF/LEF1) target genes by real-time PCR. The TCF/LEF1 target genes ( Axin2 , Cd44 , and Clcn2 ) were significantly increased in KO colonic tissues compared with WT. Data are presented as means ± SE ( n = 5–6). * P
    Figure Legend Snippet: Chloride channel protein-2 (ClC-2) knockout (KO) mice have shown disrupted adherens junctions (AJs) with nuclear localization of β-catenin in colon compared with wild-type (WT) mice. Sections of colonic tissue of ClC-2 WT and KO mice were immunolabeled for E-cadherin and β-catenin (red), and the nucleus (blue). A : ClC-2 KO colonic crypts showed dramatic disruptions of E-cadherin in upper crypts and elevated cytosolic and nuclear distribution of β-catenin. B : proximity ligation assay (PLA) showing the interaction (red dots) between E-cadherin and β-catenin in ClC-2 KO mouse colonic crypts was significantly reduced compared with ClC-2 WT. C : nuclear β-catenin was measured using ImageJ and showed significant elevations in ClC-2 KO colon ( n = 6). D : mRNA from the colonic tissues of WT and KO mice were studied for expression of T cell factor-1/lymphoid-enhancing factor-1 (TCF/LEF1) target genes by real-time PCR. The TCF/LEF1 target genes ( Axin2 , Cd44 , and Clcn2 ) were significantly increased in KO colonic tissues compared with WT. Data are presented as means ± SE ( n = 5–6). * P

    Techniques Used: Knock-Out, Mouse Assay, Immunolabeling, Proximity Ligation Assay, Expressing, Real-time Polymerase Chain Reaction

    Chloride channel protein-2 (ClC-2) has a critical function in regulation of colonic homeostasis and tumorigenicity via regulation of the stability of adherens junctions (AJs). In normal colonic mucosa, ClC-2 is associated with stability of AJ proteins to maintain colonic homeostasis. In the absence of ClC-2, colonic epithelial cells have disrupted AJ proteins E-cadherin, with internalization of β-catenin to the cytosol and subsequently the nucleus. Increased nuclear β-catenin results in alteration of colonic homeostasis and increased tumorigenicity associated with increased transcription of T cell factor-1/lymphoid-enhancing factor-1 (TCF/LEF1) target genes.
    Figure Legend Snippet: Chloride channel protein-2 (ClC-2) has a critical function in regulation of colonic homeostasis and tumorigenicity via regulation of the stability of adherens junctions (AJs). In normal colonic mucosa, ClC-2 is associated with stability of AJ proteins to maintain colonic homeostasis. In the absence of ClC-2, colonic epithelial cells have disrupted AJ proteins E-cadherin, with internalization of β-catenin to the cytosol and subsequently the nucleus. Increased nuclear β-catenin results in alteration of colonic homeostasis and increased tumorigenicity associated with increased transcription of T cell factor-1/lymphoid-enhancing factor-1 (TCF/LEF1) target genes.

    Techniques Used:

    Tight junction (TJ) proteins did not show alterations between chloride channel protein-2 (ClC-2) wild-type (WT) and knockout (KO) colon. Sections of colonic tissue of ClC-2 WT and KO mice were immunolabeled for zonula occludens-1 (ZO-1), occludin, and claudin-1 (red), and the nucleus (blue). Images were taken with an Olympus IX83 Inverted Motorized Microscope and were analyzed with cellSens software. Total RNA was extracted from colonic tissues, and real-time PCR was performed with the QuantStudio 6 Flex Real-Time PCR System. ClC-2 KO mice did not show alteration of TJ protein distribution or expression ( n = 6).
    Figure Legend Snippet: Tight junction (TJ) proteins did not show alterations between chloride channel protein-2 (ClC-2) wild-type (WT) and knockout (KO) colon. Sections of colonic tissue of ClC-2 WT and KO mice were immunolabeled for zonula occludens-1 (ZO-1), occludin, and claudin-1 (red), and the nucleus (blue). Images were taken with an Olympus IX83 Inverted Motorized Microscope and were analyzed with cellSens software. Total RNA was extracted from colonic tissues, and real-time PCR was performed with the QuantStudio 6 Flex Real-Time PCR System. ClC-2 KO mice did not show alteration of TJ protein distribution or expression ( n = 6).

    Techniques Used: Knock-Out, Mouse Assay, Immunolabeling, Microscopy, Software, Real-time Polymerase Chain Reaction, Expressing

    Colonoids from chloride channel protein-2 (ClC-2) knockout (KO) mice had altered adherens junctions (AJ) protein distribution. Colonoids from ClC-2 wild-type (WT) and KO mice were immunolabeled for E-cadherin (red), β-catenin (green), carbonic anhydrase II (CAII, green), chromogranin A (CgA, red), Muc2 (red), and the nucleus (blue). ClC-2 KO mice had evidence of disruption of E-cadherin, elevated cytosolic and nuclear distribution of β-catenin, reduced carbonic anhydrase II (CAII), and increased Muc2.
    Figure Legend Snippet: Colonoids from chloride channel protein-2 (ClC-2) knockout (KO) mice had altered adherens junctions (AJ) protein distribution. Colonoids from ClC-2 wild-type (WT) and KO mice were immunolabeled for E-cadherin (red), β-catenin (green), carbonic anhydrase II (CAII, green), chromogranin A (CgA, red), Muc2 (red), and the nucleus (blue). ClC-2 KO mice had evidence of disruption of E-cadherin, elevated cytosolic and nuclear distribution of β-catenin, reduced carbonic anhydrase II (CAII), and increased Muc2.

    Techniques Used: Knock-Out, Mouse Assay, Immunolabeling

    Chloride channel protein-2 (ClC-2) loss in intestinal crypts altered formation of colonoids. Colonoid cultures from colonic crypts of ClC-2 wild-type (WT) and knockout (KO) mice harvested to form spheroids and colonoids ( n = 3). Representative photos were taken ( A ), and quantification of spheroids ( B ) and crypts buds ( C ) was analyzed at day 6 in culture. A and B : at day 6 postculture initiation, we observed a mixed phenotype of more spheroids from ClC-2 KO colonic crypts compared with crypt-like structures in ClC-2 WT. C : the no. of buds from ClC-2 KO colonoids was significantly reduced compared with WT colonoids. D : the efficiency of colonoids from ClC-2 KO crypts (growth percentage) was not different compared with WT. E : the surface area of the colonoids was measured and showed no significant difference between two groups. Data are presented as means ± SE ( n = 3). * P
    Figure Legend Snippet: Chloride channel protein-2 (ClC-2) loss in intestinal crypts altered formation of colonoids. Colonoid cultures from colonic crypts of ClC-2 wild-type (WT) and knockout (KO) mice harvested to form spheroids and colonoids ( n = 3). Representative photos were taken ( A ), and quantification of spheroids ( B ) and crypts buds ( C ) was analyzed at day 6 in culture. A and B : at day 6 postculture initiation, we observed a mixed phenotype of more spheroids from ClC-2 KO colonic crypts compared with crypt-like structures in ClC-2 WT. C : the no. of buds from ClC-2 KO colonoids was significantly reduced compared with WT colonoids. D : the efficiency of colonoids from ClC-2 KO crypts (growth percentage) was not different compared with WT. E : the surface area of the colonoids was measured and showed no significant difference between two groups. Data are presented as means ± SE ( n = 3). * P

    Techniques Used: Knock-Out, Mouse Assay

    Gene expression profile of chloride channel protein-2 (ClC-2, clcn2 ) in colorectal cancer. Gene expression data using microarray and RNA-Seq retrieved from the National Institutes of Health Cancer Genome Atlas (TCGA) project. The expression of ClC-2 was significantly reduced compared with normal colorectal tissues.
    Figure Legend Snippet: Gene expression profile of chloride channel protein-2 (ClC-2, clcn2 ) in colorectal cancer. Gene expression data using microarray and RNA-Seq retrieved from the National Institutes of Health Cancer Genome Atlas (TCGA) project. The expression of ClC-2 was significantly reduced compared with normal colorectal tissues.

    Techniques Used: Expressing, Microarray, RNA Sequencing Assay

    Absence of chloride channel protein-2 (ClC-2) promoted the development of colitis-associated tumors. A : protocol used for the development of colitis-associated colorectal cancer (CAC) based on the coadministration of azoxymethane (AOM, 10 mg/kg ip at day 0 ) and dextran sulfate sodium (DSS, 2% in 3 cycles of 5 days each at weeks 2 , 5 , and 8 ). B : absence of ClC-2 significantly elevated tumor growth and size ( n = 8–9 for each group). ClC-2 knockout (KO) mice showed a significant elevation in the total number and size of colon tumors compared with wild-type (WT) mice. C : tumors were analyzed for dysplasia grade [low-grade dysplasia (LGD) and high-grade dysplasia (HGD)] and quantified based on percent of tumors demonstrating each grade. ClC-2 KO mice showed significantly increased HGD compared with WT mice, using a Fisher’s exact test. D and E : the proportion of proliferating cells in nontumor and tumor regions, quantified using bromodeoxyuridine (BrdU) staining, was significantly increased in the KO CAC mice compared with WT CAC mice. Data are presented as means ± SE. * P
    Figure Legend Snippet: Absence of chloride channel protein-2 (ClC-2) promoted the development of colitis-associated tumors. A : protocol used for the development of colitis-associated colorectal cancer (CAC) based on the coadministration of azoxymethane (AOM, 10 mg/kg ip at day 0 ) and dextran sulfate sodium (DSS, 2% in 3 cycles of 5 days each at weeks 2 , 5 , and 8 ). B : absence of ClC-2 significantly elevated tumor growth and size ( n = 8–9 for each group). ClC-2 knockout (KO) mice showed a significant elevation in the total number and size of colon tumors compared with wild-type (WT) mice. C : tumors were analyzed for dysplasia grade [low-grade dysplasia (LGD) and high-grade dysplasia (HGD)] and quantified based on percent of tumors demonstrating each grade. ClC-2 KO mice showed significantly increased HGD compared with WT mice, using a Fisher’s exact test. D and E : the proportion of proliferating cells in nontumor and tumor regions, quantified using bromodeoxyuridine (BrdU) staining, was significantly increased in the KO CAC mice compared with WT CAC mice. Data are presented as means ± SE. * P

    Techniques Used: Knock-Out, Mouse Assay, BrdU Staining

    Role of chloride channel protein-2 (ClC-2) in ultrastructural morphology of intercellular junctions in colon. A : to evaluate the role of ClC-2 in ultrastructural morphology of intercellular junctions, images were taken using a JEOL JEM-1200EX transmission electron microscope (TEM) and Gatan ES1000 camera system of intestines from ClC-2 wild-type (WT) and knockout (KO) mice. In small intestine ( top ), there is no dramatic morphological change between two groups compared with colon. In WT colon, tight junctions (TJs) and adherens junctions (AJs) appeared to be more electron dense and were more closely apposed compared with ClC-2 KO colon, in which TJs and AJs were less well defined and lateral epithelial membranes were tortuously folded (black arrows) ( n = 2). B : mucosa from small intestine (SI) and colon were studied for expression of ClC-2 by Western blotting. Densitometry analysis was performed to quantify expression using β-actin as a loading control. Data are presented as means ± SE ( n = 4). ** P
    Figure Legend Snippet: Role of chloride channel protein-2 (ClC-2) in ultrastructural morphology of intercellular junctions in colon. A : to evaluate the role of ClC-2 in ultrastructural morphology of intercellular junctions, images were taken using a JEOL JEM-1200EX transmission electron microscope (TEM) and Gatan ES1000 camera system of intestines from ClC-2 wild-type (WT) and knockout (KO) mice. In small intestine ( top ), there is no dramatic morphological change between two groups compared with colon. In WT colon, tight junctions (TJs) and adherens junctions (AJs) appeared to be more electron dense and were more closely apposed compared with ClC-2 KO colon, in which TJs and AJs were less well defined and lateral epithelial membranes were tortuously folded (black arrows) ( n = 2). B : mucosa from small intestine (SI) and colon were studied for expression of ClC-2 by Western blotting. Densitometry analysis was performed to quantify expression using β-actin as a loading control. Data are presented as means ± SE ( n = 4). ** P

    Techniques Used: Transmission Assay, Microscopy, Transmission Electron Microscopy, Knock-Out, Mouse Assay, Expressing, Western Blot

    Chloride channel protein-2 (ClC-2) knockout (KO) mice have altered colonic crypt differentiation. A : ClC-2 wild-type (WT) and KO mice were given bromodeoxyuridine (BrdU, ip) 24 h before being killed and analyzed ( n = 6). The ratio of BrdU-positive cells in colonic crypts was significantly increased in ClC-2 KO mice compared with WT mice. B : immunohistochemistry (IHC) for carbonic anhydrase II (CAII) as a maker of mature colonic enterocytes. The brackets indicate the region of most highly expressed CAII in the crypts, with ClC-2 KO mice having less differentiated colonic enterocytes than WT mice. C : crypt depth in hematoxylin and eosin (H E)-stained colonic cross sections, showing no significant difference between WT and ClC-2 KO mice. D : the no. of goblet cells per crypt was quantified using Alcian blue staining and showed no significant differences between two groups. E : the ratio of enterochromaffin cells per crypt was quantified using chromogranin A (CgA) staining and showed no significant difference between two groups. Data are presented as means ± SE ( n = 6). *** P
    Figure Legend Snippet: Chloride channel protein-2 (ClC-2) knockout (KO) mice have altered colonic crypt differentiation. A : ClC-2 wild-type (WT) and KO mice were given bromodeoxyuridine (BrdU, ip) 24 h before being killed and analyzed ( n = 6). The ratio of BrdU-positive cells in colonic crypts was significantly increased in ClC-2 KO mice compared with WT mice. B : immunohistochemistry (IHC) for carbonic anhydrase II (CAII) as a maker of mature colonic enterocytes. The brackets indicate the region of most highly expressed CAII in the crypts, with ClC-2 KO mice having less differentiated colonic enterocytes than WT mice. C : crypt depth in hematoxylin and eosin (H E)-stained colonic cross sections, showing no significant difference between WT and ClC-2 KO mice. D : the no. of goblet cells per crypt was quantified using Alcian blue staining and showed no significant differences between two groups. E : the ratio of enterochromaffin cells per crypt was quantified using chromogranin A (CgA) staining and showed no significant difference between two groups. Data are presented as means ± SE ( n = 6). *** P

    Techniques Used: Knock-Out, Mouse Assay, Immunohistochemistry, Staining

    3) Product Images from "Parallel intermediate conductance K+ and Cl− channel activity mediates electroneutral K+ exit across basolateral membranes in rat distal colon"

    Article Title: Parallel intermediate conductance K+ and Cl− channel activity mediates electroneutral K+ exit across basolateral membranes in rat distal colon

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    doi: 10.1152/ajpgi.00011.2020

    Localization of CLC2- and Na + -K + -ATPase α-subunit (NaKα)-like proteins in rat distal colon. A and A’ : CLC2-like proteins (green) localized predominantly in surface cells (arrows), while minimally expressed in crypt regions (asterisks). B and B’ : NaKα-like proteins (red) localized to both surface (arrows) and crypt (asterisks) cells. C and C’ : nuclei (blue) stained with Hoechst 33342. D : composite of A , B , and C . D’ : composite of A’ , B’ , and C’ . Higher magnification of the boxed region from D and D’ presented in E and E’ , respectively. E : CLC2 and NaKα colocalized (orange) in both lateral (arrows) and basement (arrowheads) membranes of surface cells. E’ : in crypt cells, NaKα (arrow) and CLC2 (arrowhead) localized in the basolateral membranes of crypts. Identical results obtained in different specimens obtained from three different experiments with three different male and female (not shown) rat distal colons.
    Figure Legend Snippet: Localization of CLC2- and Na + -K + -ATPase α-subunit (NaKα)-like proteins in rat distal colon. A and A’ : CLC2-like proteins (green) localized predominantly in surface cells (arrows), while minimally expressed in crypt regions (asterisks). B and B’ : NaKα-like proteins (red) localized to both surface (arrows) and crypt (asterisks) cells. C and C’ : nuclei (blue) stained with Hoechst 33342. D : composite of A , B , and C . D’ : composite of A’ , B’ , and C’ . Higher magnification of the boxed region from D and D’ presented in E and E’ , respectively. E : CLC2 and NaKα colocalized (orange) in both lateral (arrows) and basement (arrowheads) membranes of surface cells. E’ : in crypt cells, NaKα (arrow) and CLC2 (arrowhead) localized in the basolateral membranes of crypts. Identical results obtained in different specimens obtained from three different experiments with three different male and female (not shown) rat distal colons.

    Techniques Used: Staining

    Related Articles

    Binding Assay:

    Article Title: Functional characterization of a ClC-2-like Cl(-) conductance in surface epithelial cells of rat rectal colon.
    Article Snippet: .. ClC-2, a member of the voltage-gated Cl(-) channel family, is expressed in the distal colonic surface epithelial cells of various species, but its functional significance remains unclear.. Here, by means of electrophysiological and molecular biological techniques, we have identified and characterized a ClC-2-like conductance naturally expressed by surface epithelial cells acutely dissociated from rectal colon of rats fed a standard diet. ..

    Incubation:

    Article Title: Functional characterization of a ClC-2-like Cl(-) conductance in surface epithelial cells of rat rectal colon.
    Article Snippet: .. ClC-2, a member of the voltage-gated Cl(-) channel family, is expressed in the distal colonic surface epithelial cells of various species, but its functional significance remains unclear.. Here, by means of electrophysiological and molecular biological techniques, we have identified and characterized a ClC-2-like conductance naturally expressed by surface epithelial cells acutely dissociated from rectal colon of rats fed a standard diet. ..

    Article Title: Parallel intermediate conductance K+ and Cl− channel activity mediates electroneutral K+ exit across basolateral membranes in rat distal colon
    Article Snippet: .. Sections (10 μm) were placed on charged slides and exposed to 5% goat serum for 2 h. After the sections were rinsed with PBS containing 0.1% Triton X-100 and 0.1% toluene (PBS-Tween) for 10 min, they were incubated overnight with either a primary antibody to Na+ -K+-ATPase α-subunit (NaKα1; 1:1,000 dilution, kindly provided by Dr. Michael Kashgarian, Yale School of Medicine, New Haven, CT) ( , ), or anti-CLC2 antibody (1:500 dilution; cat. no. ACL002; Alomone Laboratories, Jerusalem, Israel) diluted with 5% goat serum. ..

    Article Title: Lubiprostone activates Cl- secretion via cAMP signaling and increases membrane CFTR in the human colon carcinoma cell line, T84.
    Article Snippet: .. Lubiprostone, used clinically (b.i.d.) to treat constipation, has been reported to increase transepithelial Cl(-) transport in T84 cells by activating ClC-2 channels.. To identify the underlying signaling pathway, we explored the effects of short-term and overnight lubiprostone treatment on second messenger signaling and Cl(-) transport. ..

    Article Title: ClC-2 contributes to tonic inhibition mediated by α5 subunit-containing GABA(A) receptor in experimental temporal lobe epilepsy.
    Article Snippet: .. Temporal lobe epilepsy (TLE), characterized by spontaneous recurrent seizures, learning and memory impairments is associated with neurodegeneration, abnormal reorganization of the circuitry and loss of functional inhibition in hippocampus.. In adult hippocampus, the GABAergic cells mediate the major inhibitory function of the principal neurons, promoting the Cl(-) entry through the GABA(A) receptor, whether through phasic (synaptic) or tonic (extrasynaptic) conductance. ..

    Staining:

    Article Title: Molecular identification of HSPA8 as an accessory protein of a hyperpolarization-activated chloride channel from rat pulmonary vein cardiomyocytes
    Article Snippet: .. Cells were then stained with antibodies against ryanodine receptor (RyR; Abcam, Cambridge, UK; catalog no. ab2827), Na/K pump (Abcam, catalog no. ab76020), CLCN2 (Santa Cruz Biotechnology, Dallas, TX, catalog no. sc-377284; Alomone Labs, Jerusalem, Israel, catalog no. ACL-002), and HSPA8 (Abcam; catalog no. ab2788). ..

    Blocking Assay:

    Article Title: ClC-2 contributes to tonic inhibition mediated by α5 subunit-containing GABA(A) receptor in experimental temporal lobe epilepsy.
    Article Snippet: .. Temporal lobe epilepsy (TLE), characterized by spontaneous recurrent seizures, learning and memory impairments is associated with neurodegeneration, abnormal reorganization of the circuitry and loss of functional inhibition in hippocampus.. In adult hippocampus, the GABAergic cells mediate the major inhibitory function of the principal neurons, promoting the Cl(-) entry through the GABA(A) receptor, whether through phasic (synaptic) or tonic (extrasynaptic) conductance. ..

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    Alomone Labs mouse clc 2
    Functional expression of the <t>ClC-2</t> channel in submandibular granular duct cells. A : differential interference contrast images of single granular duct and acinar cells freshly isolated from male mouse submandibular glands. Scale bar = 10 μm
    Mouse Clc 2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse clc 2/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse clc 2 - by Bioz Stars, 2021-09
    93/100 stars
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    Image Search Results


    Functional expression of the ClC-2 channel in submandibular granular duct cells. A : differential interference contrast images of single granular duct and acinar cells freshly isolated from male mouse submandibular glands. Scale bar = 10 μm

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Clcn2 encodes the hyperpolarization-activated chloride channel in the ducts of mouse salivary glands

    doi: 10.1152/ajpgi.90384.2008

    Figure Lengend Snippet: Functional expression of the ClC-2 channel in submandibular granular duct cells. A : differential interference contrast images of single granular duct and acinar cells freshly isolated from male mouse submandibular glands. Scale bar = 10 μm

    Article Snippet: This antibody recognizes the COOH-terminus epitope RSRHGLPREGTPSDSDDKC of rat ClC-2 that is identical to mouse ClC-2 (ACL-002, Alomone Labs, Jerusalem, Israel).

    Techniques: Functional Assay, Expressing, Isolation

    ClC-2 distribution in mouse submandibular and parotid glands. A : immunoperoxidase labeling for ClC-2 in the submandibular gland of a Clcn2 +/+ male mouse. AC, acinar cells; GD, granular duct; SD, striated duct; ED, excretory duct. Inset,

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Clcn2 encodes the hyperpolarization-activated chloride channel in the ducts of mouse salivary glands

    doi: 10.1152/ajpgi.90384.2008

    Figure Lengend Snippet: ClC-2 distribution in mouse submandibular and parotid glands. A : immunoperoxidase labeling for ClC-2 in the submandibular gland of a Clcn2 +/+ male mouse. AC, acinar cells; GD, granular duct; SD, striated duct; ED, excretory duct. Inset,

    Article Snippet: This antibody recognizes the COOH-terminus epitope RSRHGLPREGTPSDSDDKC of rat ClC-2 that is identical to mouse ClC-2 (ACL-002, Alomone Labs, Jerusalem, Israel).

    Techniques: Labeling

    Chloride channel protein-2 (ClC-2) knockout (KO) mice have shown disrupted adherens junctions (AJs) with nuclear localization of β-catenin in colon compared with wild-type (WT) mice. Sections of colonic tissue of ClC-2 WT and KO mice were immunolabeled for E-cadherin and β-catenin (red), and the nucleus (blue). A : ClC-2 KO colonic crypts showed dramatic disruptions of E-cadherin in upper crypts and elevated cytosolic and nuclear distribution of β-catenin. B : proximity ligation assay (PLA) showing the interaction (red dots) between E-cadherin and β-catenin in ClC-2 KO mouse colonic crypts was significantly reduced compared with ClC-2 WT. C : nuclear β-catenin was measured using ImageJ and showed significant elevations in ClC-2 KO colon ( n = 6). D : mRNA from the colonic tissues of WT and KO mice were studied for expression of T cell factor-1/lymphoid-enhancing factor-1 (TCF/LEF1) target genes by real-time PCR. The TCF/LEF1 target genes ( Axin2 , Cd44 , and Clcn2 ) were significantly increased in KO colonic tissues compared with WT. Data are presented as means ± SE ( n = 5–6). * P

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Epithelial Biology and Secretion: Knockout of ClC-2 reveals critical functions of adherens junctions in colonic homeostasis and tumorigenicity

    doi: 10.1152/ajpgi.00087.2018

    Figure Lengend Snippet: Chloride channel protein-2 (ClC-2) knockout (KO) mice have shown disrupted adherens junctions (AJs) with nuclear localization of β-catenin in colon compared with wild-type (WT) mice. Sections of colonic tissue of ClC-2 WT and KO mice were immunolabeled for E-cadherin and β-catenin (red), and the nucleus (blue). A : ClC-2 KO colonic crypts showed dramatic disruptions of E-cadherin in upper crypts and elevated cytosolic and nuclear distribution of β-catenin. B : proximity ligation assay (PLA) showing the interaction (red dots) between E-cadherin and β-catenin in ClC-2 KO mouse colonic crypts was significantly reduced compared with ClC-2 WT. C : nuclear β-catenin was measured using ImageJ and showed significant elevations in ClC-2 KO colon ( n = 6). D : mRNA from the colonic tissues of WT and KO mice were studied for expression of T cell factor-1/lymphoid-enhancing factor-1 (TCF/LEF1) target genes by real-time PCR. The TCF/LEF1 target genes ( Axin2 , Cd44 , and Clcn2 ) were significantly increased in KO colonic tissues compared with WT. Data are presented as means ± SE ( n = 5–6). * P

    Article Snippet: After protein transfer to a polyvinylidene difluoride membrane, the membranes were probed using anti-ClC-2 (1:200, ACL-002; Alomone Laboratories, Jerusalem, Israel) and anti-β-actin (1:20,000, ab8227; Abcam) antibodies.

    Techniques: Knock-Out, Mouse Assay, Immunolabeling, Proximity Ligation Assay, Expressing, Real-time Polymerase Chain Reaction

    Chloride channel protein-2 (ClC-2) has a critical function in regulation of colonic homeostasis and tumorigenicity via regulation of the stability of adherens junctions (AJs). In normal colonic mucosa, ClC-2 is associated with stability of AJ proteins to maintain colonic homeostasis. In the absence of ClC-2, colonic epithelial cells have disrupted AJ proteins E-cadherin, with internalization of β-catenin to the cytosol and subsequently the nucleus. Increased nuclear β-catenin results in alteration of colonic homeostasis and increased tumorigenicity associated with increased transcription of T cell factor-1/lymphoid-enhancing factor-1 (TCF/LEF1) target genes.

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Epithelial Biology and Secretion: Knockout of ClC-2 reveals critical functions of adherens junctions in colonic homeostasis and tumorigenicity

    doi: 10.1152/ajpgi.00087.2018

    Figure Lengend Snippet: Chloride channel protein-2 (ClC-2) has a critical function in regulation of colonic homeostasis and tumorigenicity via regulation of the stability of adherens junctions (AJs). In normal colonic mucosa, ClC-2 is associated with stability of AJ proteins to maintain colonic homeostasis. In the absence of ClC-2, colonic epithelial cells have disrupted AJ proteins E-cadherin, with internalization of β-catenin to the cytosol and subsequently the nucleus. Increased nuclear β-catenin results in alteration of colonic homeostasis and increased tumorigenicity associated with increased transcription of T cell factor-1/lymphoid-enhancing factor-1 (TCF/LEF1) target genes.

    Article Snippet: After protein transfer to a polyvinylidene difluoride membrane, the membranes were probed using anti-ClC-2 (1:200, ACL-002; Alomone Laboratories, Jerusalem, Israel) and anti-β-actin (1:20,000, ab8227; Abcam) antibodies.

    Techniques:

    Tight junction (TJ) proteins did not show alterations between chloride channel protein-2 (ClC-2) wild-type (WT) and knockout (KO) colon. Sections of colonic tissue of ClC-2 WT and KO mice were immunolabeled for zonula occludens-1 (ZO-1), occludin, and claudin-1 (red), and the nucleus (blue). Images were taken with an Olympus IX83 Inverted Motorized Microscope and were analyzed with cellSens software. Total RNA was extracted from colonic tissues, and real-time PCR was performed with the QuantStudio 6 Flex Real-Time PCR System. ClC-2 KO mice did not show alteration of TJ protein distribution or expression ( n = 6).

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Epithelial Biology and Secretion: Knockout of ClC-2 reveals critical functions of adherens junctions in colonic homeostasis and tumorigenicity

    doi: 10.1152/ajpgi.00087.2018

    Figure Lengend Snippet: Tight junction (TJ) proteins did not show alterations between chloride channel protein-2 (ClC-2) wild-type (WT) and knockout (KO) colon. Sections of colonic tissue of ClC-2 WT and KO mice were immunolabeled for zonula occludens-1 (ZO-1), occludin, and claudin-1 (red), and the nucleus (blue). Images were taken with an Olympus IX83 Inverted Motorized Microscope and were analyzed with cellSens software. Total RNA was extracted from colonic tissues, and real-time PCR was performed with the QuantStudio 6 Flex Real-Time PCR System. ClC-2 KO mice did not show alteration of TJ protein distribution or expression ( n = 6).

    Article Snippet: After protein transfer to a polyvinylidene difluoride membrane, the membranes were probed using anti-ClC-2 (1:200, ACL-002; Alomone Laboratories, Jerusalem, Israel) and anti-β-actin (1:20,000, ab8227; Abcam) antibodies.

    Techniques: Knock-Out, Mouse Assay, Immunolabeling, Microscopy, Software, Real-time Polymerase Chain Reaction, Expressing

    Colonoids from chloride channel protein-2 (ClC-2) knockout (KO) mice had altered adherens junctions (AJ) protein distribution. Colonoids from ClC-2 wild-type (WT) and KO mice were immunolabeled for E-cadherin (red), β-catenin (green), carbonic anhydrase II (CAII, green), chromogranin A (CgA, red), Muc2 (red), and the nucleus (blue). ClC-2 KO mice had evidence of disruption of E-cadherin, elevated cytosolic and nuclear distribution of β-catenin, reduced carbonic anhydrase II (CAII), and increased Muc2.

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Epithelial Biology and Secretion: Knockout of ClC-2 reveals critical functions of adherens junctions in colonic homeostasis and tumorigenicity

    doi: 10.1152/ajpgi.00087.2018

    Figure Lengend Snippet: Colonoids from chloride channel protein-2 (ClC-2) knockout (KO) mice had altered adherens junctions (AJ) protein distribution. Colonoids from ClC-2 wild-type (WT) and KO mice were immunolabeled for E-cadherin (red), β-catenin (green), carbonic anhydrase II (CAII, green), chromogranin A (CgA, red), Muc2 (red), and the nucleus (blue). ClC-2 KO mice had evidence of disruption of E-cadherin, elevated cytosolic and nuclear distribution of β-catenin, reduced carbonic anhydrase II (CAII), and increased Muc2.

    Article Snippet: After protein transfer to a polyvinylidene difluoride membrane, the membranes were probed using anti-ClC-2 (1:200, ACL-002; Alomone Laboratories, Jerusalem, Israel) and anti-β-actin (1:20,000, ab8227; Abcam) antibodies.

    Techniques: Knock-Out, Mouse Assay, Immunolabeling

    Chloride channel protein-2 (ClC-2) loss in intestinal crypts altered formation of colonoids. Colonoid cultures from colonic crypts of ClC-2 wild-type (WT) and knockout (KO) mice harvested to form spheroids and colonoids ( n = 3). Representative photos were taken ( A ), and quantification of spheroids ( B ) and crypts buds ( C ) was analyzed at day 6 in culture. A and B : at day 6 postculture initiation, we observed a mixed phenotype of more spheroids from ClC-2 KO colonic crypts compared with crypt-like structures in ClC-2 WT. C : the no. of buds from ClC-2 KO colonoids was significantly reduced compared with WT colonoids. D : the efficiency of colonoids from ClC-2 KO crypts (growth percentage) was not different compared with WT. E : the surface area of the colonoids was measured and showed no significant difference between two groups. Data are presented as means ± SE ( n = 3). * P

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Epithelial Biology and Secretion: Knockout of ClC-2 reveals critical functions of adherens junctions in colonic homeostasis and tumorigenicity

    doi: 10.1152/ajpgi.00087.2018

    Figure Lengend Snippet: Chloride channel protein-2 (ClC-2) loss in intestinal crypts altered formation of colonoids. Colonoid cultures from colonic crypts of ClC-2 wild-type (WT) and knockout (KO) mice harvested to form spheroids and colonoids ( n = 3). Representative photos were taken ( A ), and quantification of spheroids ( B ) and crypts buds ( C ) was analyzed at day 6 in culture. A and B : at day 6 postculture initiation, we observed a mixed phenotype of more spheroids from ClC-2 KO colonic crypts compared with crypt-like structures in ClC-2 WT. C : the no. of buds from ClC-2 KO colonoids was significantly reduced compared with WT colonoids. D : the efficiency of colonoids from ClC-2 KO crypts (growth percentage) was not different compared with WT. E : the surface area of the colonoids was measured and showed no significant difference between two groups. Data are presented as means ± SE ( n = 3). * P

    Article Snippet: After protein transfer to a polyvinylidene difluoride membrane, the membranes were probed using anti-ClC-2 (1:200, ACL-002; Alomone Laboratories, Jerusalem, Israel) and anti-β-actin (1:20,000, ab8227; Abcam) antibodies.

    Techniques: Knock-Out, Mouse Assay

    Gene expression profile of chloride channel protein-2 (ClC-2, clcn2 ) in colorectal cancer. Gene expression data using microarray and RNA-Seq retrieved from the National Institutes of Health Cancer Genome Atlas (TCGA) project. The expression of ClC-2 was significantly reduced compared with normal colorectal tissues.

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Epithelial Biology and Secretion: Knockout of ClC-2 reveals critical functions of adherens junctions in colonic homeostasis and tumorigenicity

    doi: 10.1152/ajpgi.00087.2018

    Figure Lengend Snippet: Gene expression profile of chloride channel protein-2 (ClC-2, clcn2 ) in colorectal cancer. Gene expression data using microarray and RNA-Seq retrieved from the National Institutes of Health Cancer Genome Atlas (TCGA) project. The expression of ClC-2 was significantly reduced compared with normal colorectal tissues.

    Article Snippet: After protein transfer to a polyvinylidene difluoride membrane, the membranes were probed using anti-ClC-2 (1:200, ACL-002; Alomone Laboratories, Jerusalem, Israel) and anti-β-actin (1:20,000, ab8227; Abcam) antibodies.

    Techniques: Expressing, Microarray, RNA Sequencing Assay

    Absence of chloride channel protein-2 (ClC-2) promoted the development of colitis-associated tumors. A : protocol used for the development of colitis-associated colorectal cancer (CAC) based on the coadministration of azoxymethane (AOM, 10 mg/kg ip at day 0 ) and dextran sulfate sodium (DSS, 2% in 3 cycles of 5 days each at weeks 2 , 5 , and 8 ). B : absence of ClC-2 significantly elevated tumor growth and size ( n = 8–9 for each group). ClC-2 knockout (KO) mice showed a significant elevation in the total number and size of colon tumors compared with wild-type (WT) mice. C : tumors were analyzed for dysplasia grade [low-grade dysplasia (LGD) and high-grade dysplasia (HGD)] and quantified based on percent of tumors demonstrating each grade. ClC-2 KO mice showed significantly increased HGD compared with WT mice, using a Fisher’s exact test. D and E : the proportion of proliferating cells in nontumor and tumor regions, quantified using bromodeoxyuridine (BrdU) staining, was significantly increased in the KO CAC mice compared with WT CAC mice. Data are presented as means ± SE. * P

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Epithelial Biology and Secretion: Knockout of ClC-2 reveals critical functions of adherens junctions in colonic homeostasis and tumorigenicity

    doi: 10.1152/ajpgi.00087.2018

    Figure Lengend Snippet: Absence of chloride channel protein-2 (ClC-2) promoted the development of colitis-associated tumors. A : protocol used for the development of colitis-associated colorectal cancer (CAC) based on the coadministration of azoxymethane (AOM, 10 mg/kg ip at day 0 ) and dextran sulfate sodium (DSS, 2% in 3 cycles of 5 days each at weeks 2 , 5 , and 8 ). B : absence of ClC-2 significantly elevated tumor growth and size ( n = 8–9 for each group). ClC-2 knockout (KO) mice showed a significant elevation in the total number and size of colon tumors compared with wild-type (WT) mice. C : tumors were analyzed for dysplasia grade [low-grade dysplasia (LGD) and high-grade dysplasia (HGD)] and quantified based on percent of tumors demonstrating each grade. ClC-2 KO mice showed significantly increased HGD compared with WT mice, using a Fisher’s exact test. D and E : the proportion of proliferating cells in nontumor and tumor regions, quantified using bromodeoxyuridine (BrdU) staining, was significantly increased in the KO CAC mice compared with WT CAC mice. Data are presented as means ± SE. * P

    Article Snippet: After protein transfer to a polyvinylidene difluoride membrane, the membranes were probed using anti-ClC-2 (1:200, ACL-002; Alomone Laboratories, Jerusalem, Israel) and anti-β-actin (1:20,000, ab8227; Abcam) antibodies.

    Techniques: Knock-Out, Mouse Assay, BrdU Staining

    Role of chloride channel protein-2 (ClC-2) in ultrastructural morphology of intercellular junctions in colon. A : to evaluate the role of ClC-2 in ultrastructural morphology of intercellular junctions, images were taken using a JEOL JEM-1200EX transmission electron microscope (TEM) and Gatan ES1000 camera system of intestines from ClC-2 wild-type (WT) and knockout (KO) mice. In small intestine ( top ), there is no dramatic morphological change between two groups compared with colon. In WT colon, tight junctions (TJs) and adherens junctions (AJs) appeared to be more electron dense and were more closely apposed compared with ClC-2 KO colon, in which TJs and AJs were less well defined and lateral epithelial membranes were tortuously folded (black arrows) ( n = 2). B : mucosa from small intestine (SI) and colon were studied for expression of ClC-2 by Western blotting. Densitometry analysis was performed to quantify expression using β-actin as a loading control. Data are presented as means ± SE ( n = 4). ** P

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Epithelial Biology and Secretion: Knockout of ClC-2 reveals critical functions of adherens junctions in colonic homeostasis and tumorigenicity

    doi: 10.1152/ajpgi.00087.2018

    Figure Lengend Snippet: Role of chloride channel protein-2 (ClC-2) in ultrastructural morphology of intercellular junctions in colon. A : to evaluate the role of ClC-2 in ultrastructural morphology of intercellular junctions, images were taken using a JEOL JEM-1200EX transmission electron microscope (TEM) and Gatan ES1000 camera system of intestines from ClC-2 wild-type (WT) and knockout (KO) mice. In small intestine ( top ), there is no dramatic morphological change between two groups compared with colon. In WT colon, tight junctions (TJs) and adherens junctions (AJs) appeared to be more electron dense and were more closely apposed compared with ClC-2 KO colon, in which TJs and AJs were less well defined and lateral epithelial membranes were tortuously folded (black arrows) ( n = 2). B : mucosa from small intestine (SI) and colon were studied for expression of ClC-2 by Western blotting. Densitometry analysis was performed to quantify expression using β-actin as a loading control. Data are presented as means ± SE ( n = 4). ** P

    Article Snippet: After protein transfer to a polyvinylidene difluoride membrane, the membranes were probed using anti-ClC-2 (1:200, ACL-002; Alomone Laboratories, Jerusalem, Israel) and anti-β-actin (1:20,000, ab8227; Abcam) antibodies.

    Techniques: Transmission Assay, Microscopy, Transmission Electron Microscopy, Knock-Out, Mouse Assay, Expressing, Western Blot

    Chloride channel protein-2 (ClC-2) knockout (KO) mice have altered colonic crypt differentiation. A : ClC-2 wild-type (WT) and KO mice were given bromodeoxyuridine (BrdU, ip) 24 h before being killed and analyzed ( n = 6). The ratio of BrdU-positive cells in colonic crypts was significantly increased in ClC-2 KO mice compared with WT mice. B : immunohistochemistry (IHC) for carbonic anhydrase II (CAII) as a maker of mature colonic enterocytes. The brackets indicate the region of most highly expressed CAII in the crypts, with ClC-2 KO mice having less differentiated colonic enterocytes than WT mice. C : crypt depth in hematoxylin and eosin (H E)-stained colonic cross sections, showing no significant difference between WT and ClC-2 KO mice. D : the no. of goblet cells per crypt was quantified using Alcian blue staining and showed no significant differences between two groups. E : the ratio of enterochromaffin cells per crypt was quantified using chromogranin A (CgA) staining and showed no significant difference between two groups. Data are presented as means ± SE ( n = 6). *** P

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Epithelial Biology and Secretion: Knockout of ClC-2 reveals critical functions of adherens junctions in colonic homeostasis and tumorigenicity

    doi: 10.1152/ajpgi.00087.2018

    Figure Lengend Snippet: Chloride channel protein-2 (ClC-2) knockout (KO) mice have altered colonic crypt differentiation. A : ClC-2 wild-type (WT) and KO mice were given bromodeoxyuridine (BrdU, ip) 24 h before being killed and analyzed ( n = 6). The ratio of BrdU-positive cells in colonic crypts was significantly increased in ClC-2 KO mice compared with WT mice. B : immunohistochemistry (IHC) for carbonic anhydrase II (CAII) as a maker of mature colonic enterocytes. The brackets indicate the region of most highly expressed CAII in the crypts, with ClC-2 KO mice having less differentiated colonic enterocytes than WT mice. C : crypt depth in hematoxylin and eosin (H E)-stained colonic cross sections, showing no significant difference between WT and ClC-2 KO mice. D : the no. of goblet cells per crypt was quantified using Alcian blue staining and showed no significant differences between two groups. E : the ratio of enterochromaffin cells per crypt was quantified using chromogranin A (CgA) staining and showed no significant difference between two groups. Data are presented as means ± SE ( n = 6). *** P

    Article Snippet: After protein transfer to a polyvinylidene difluoride membrane, the membranes were probed using anti-ClC-2 (1:200, ACL-002; Alomone Laboratories, Jerusalem, Israel) and anti-β-actin (1:20,000, ab8227; Abcam) antibodies.

    Techniques: Knock-Out, Mouse Assay, Immunohistochemistry, Staining

    Localization of CLC2- and Na + -K + -ATPase α-subunit (NaKα)-like proteins in rat distal colon. A and A’ : CLC2-like proteins (green) localized predominantly in surface cells (arrows), while minimally expressed in crypt regions (asterisks). B and B’ : NaKα-like proteins (red) localized to both surface (arrows) and crypt (asterisks) cells. C and C’ : nuclei (blue) stained with Hoechst 33342. D : composite of A , B , and C . D’ : composite of A’ , B’ , and C’ . Higher magnification of the boxed region from D and D’ presented in E and E’ , respectively. E : CLC2 and NaKα colocalized (orange) in both lateral (arrows) and basement (arrowheads) membranes of surface cells. E’ : in crypt cells, NaKα (arrow) and CLC2 (arrowhead) localized in the basolateral membranes of crypts. Identical results obtained in different specimens obtained from three different experiments with three different male and female (not shown) rat distal colons.

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Parallel intermediate conductance K+ and Cl− channel activity mediates electroneutral K+ exit across basolateral membranes in rat distal colon

    doi: 10.1152/ajpgi.00011.2020

    Figure Lengend Snippet: Localization of CLC2- and Na + -K + -ATPase α-subunit (NaKα)-like proteins in rat distal colon. A and A’ : CLC2-like proteins (green) localized predominantly in surface cells (arrows), while minimally expressed in crypt regions (asterisks). B and B’ : NaKα-like proteins (red) localized to both surface (arrows) and crypt (asterisks) cells. C and C’ : nuclei (blue) stained with Hoechst 33342. D : composite of A , B , and C . D’ : composite of A’ , B’ , and C’ . Higher magnification of the boxed region from D and D’ presented in E and E’ , respectively. E : CLC2 and NaKα colocalized (orange) in both lateral (arrows) and basement (arrowheads) membranes of surface cells. E’ : in crypt cells, NaKα (arrow) and CLC2 (arrowhead) localized in the basolateral membranes of crypts. Identical results obtained in different specimens obtained from three different experiments with three different male and female (not shown) rat distal colons.

    Article Snippet: Sections (10 μm) were placed on charged slides and exposed to 5% goat serum for 2 h. After the sections were rinsed with PBS containing 0.1% Triton X-100 and 0.1% toluene (PBS-Tween) for 10 min, they were incubated overnight with either a primary antibody to Na+ -K+-ATPase α-subunit (NaKα1; 1:1,000 dilution, kindly provided by Dr. Michael Kashgarian, Yale School of Medicine, New Haven, CT) ( , ), or anti-CLC2 antibody (1:500 dilution; cat. no. ACL002; Alomone Laboratories, Jerusalem, Israel) diluted with 5% goat serum.

    Techniques: Staining