Anti-Cytokeratin Pan (mouse monoclonal)

Species Reactivity	Key Applications	Host	Format	Antibody Type
Ma	IHC	M	Purified	Monoclonal Antibody

Species Reactivity

Key Applications

Host

Format

Antibody Type

IHC

Purified

Monoclonal Antibody

Description
Catalogue Number	MAB3406-40UG
Brand Family	Chemicon®
Trade Name	Chemicon
Description	Anti-Cytokeratin Pan Antibody, clone Lu5

Description

Catalogue Number

MAB3406-40UG

Brand Family

Chemicon®

Trade Name

Chemicon

Description

Anti-Cytokeratin Pan Antibody, clone Lu5

Product Information
Format	Purified
Presentation	Liquid. Buffer = 0.02 M Phosphate buffer, 0.25M NaCl with 0.1% sodium azide.

Product Information

Format

Purified

Presentation

Liquid. Buffer = 0.02 M Phosphate buffer, 0.25M NaCl with 0.1% sodium azide.

Applications
Application	This Anti-Cytokeratin Pan Antibody, clone Lu5 is validated for use in IH for the detection of Cytokeratin Pan.
Key Applications	Immunohistochemistry
Application Notes	Immunohistochemistry: 10-20 μg/mL Optimal working dilutions must be determined by end user. Tumor Characterization For tumor characterization a two-step experimental assay is recommended: 1. Characterization of the tissue origin of a tumor with the aid of anti-cytokeratin pan, anti-desmin, anti-glial fibrillary acidic protein, anti-neurofilament and anti-vimentin. 2. Characterization of an epithelial tumor using antibodies to cytokeratin classes which are characteristic for different tissues, e.g. anti-cytokeratin No. 7, anti-cytokeratin No. 18. Sample material: Normal tissue, tumor tissue following surgery or up to 24 h after autopsy, frozen sections or formaldehyde-fixed paraffin embedded sections, Detection method Immunohistochemical analysis (detection of the primary antibody e. g. by anti-mouse Ig-peroxidase or anti-mouse Ig-FITC). Procedure: Ideal frozen sections are obtained from shock-frozen tissue samples. The frozen sections are air-dried and then fixed with acetone for 5-10 min at -15 to -25°C. Excess acetone is allowed to evaporate at 15-25°C. Material fixed in alcohol can also be used. Cytocentrifuge preparations of single cells or cell smears are also fixed in acetone. These preparations should, however, not be air dried, the excess acetone is removed by briefly washing in phosphate-buffered saline (PBS). Further treatment as follows: • Overlay the preparation with 10 - 20 μL antibody solution and incubate in a humid chamber for 30 - 60 min at 15-25°C. • Dip the slide briefly in PBS and then wash in PBS 3 times for 3 min each (using a fresh PBS bath in each case). • Wipe the margins of the preparation dry, overlay the preparation with 10 - 20 μL of a solution of anti-mouse lg-FITC or anti-mouse Ig-peroxidase and allow to incubate in a humid chamber for 30 min at 15-25°C. • Wash the slide in PBS as described above. The preparation must not be allowed to dry out during any of the steps. lf using an indirect immunofluorescence technique, the preparation should be overlaid with a suitable embedding medium (e. g. Mowiol, Hoechst) and examined under the fluorescence microscope. lf a POD-conjugate is used as the secondary antibody, the preparation should be overlaid with a substrate solution (see below) and incubated at 15-25°C until a clearly visible redbrown color develops. A negative control (e.g. only the labeled secondary antibody) should remain unchanged in color during this incubation period. Subsequently, the substrate is washed off with PBS and the preparation is stained with hemalum stain for about 1 min. The hemalum solution is washed off with PBS, the preparation is embedded and examined. Substrate solutions: Aminoethyl-carbazole: Dissolve 2 mg 3-amino-9-ethylcarbazole with 1.2 mL dimethylsulfoxide and add 28.8 mL Tris-HCI, 0.05 M; pH 7.3; and 20 μL 3% H 2 O 2 , (w/v). Prepare solution freshly each day. Diaminobenzidine: Dissolve 25 mg 3, 3'-diaminobenzidine with 50 mL Tris-HCI, 0.05 M; pH 7.3; and add 40 μL 3% H2O2 , (w/v). Prepare solution freshly each day. Note The antibody solutions have to be absolutely free of precipitate! lf necessary, centrifuge the solutions at high speed prior to use.

Applications

Application

This Anti-Cytokeratin Pan Antibody, clone Lu5 is validated for use in IH for the detection of Cytokeratin Pan.

Key Applications

Immunohistochemistry

Application Notes

Immunohistochemistry: 10-20 μg/mL Optimal working dilutions must be determined by end user.

Tumor Characterization

For tumor characterization a two-step experimental assay is recommended:

1. Characterization of the tissue origin of a tumor with the aid of anti-cytokeratin pan, anti-desmin, anti-glial fibrillary acidic protein, anti-neurofilament and anti-vimentin.

2. Characterization of an epithelial tumor using antibodies to cytokeratin classes which are characteristic for different tissues, e.g. anti-cytokeratin No. 7, anti-cytokeratin No. 18. Sample material: Normal tissue, tumor tissue following surgery or up to 24 h after autopsy, frozen sections or formaldehyde-fixed paraffin embedded sections,

Detection method

Immunohistochemical analysis (detection of the primary antibody e. g. by anti-mouse Ig-peroxidase or anti-mouse Ig-FITC).

Procedure:

Ideal frozen sections are obtained from shock-frozen tissue samples. The frozen sections are air-dried and then fixed with acetone for 5-10 min at -15 to -25°C. Excess acetone is allowed to evaporate at 15-25°C. Material fixed in alcohol can also be used.

Cytocentrifuge preparations of single cells or cell smears are also fixed in acetone. These preparations should, however, not be air dried, the excess acetone is removed by briefly washing in phosphate-buffered saline (PBS). Further treatment as follows:

• Overlay the preparation with 10 - 20 μL antibody solution and incubate in a humid chamber for 30 - 60 min at 15-25°C.

• Dip the slide briefly in PBS and then wash in PBS 3 times for 3 min each (using a fresh PBS bath in each case).

• Wipe the margins of the preparation dry, overlay the preparation with 10 - 20 μL of a solution of anti-mouse lg-FITC or anti-mouse Ig-peroxidase and allow to incubate in a humid chamber for 30 min at 15-25°C.

• Wash the slide in PBS as described above.

The preparation must not be allowed to dry out during any of the steps. lf using an indirect immunofluorescence technique, the preparation should be overlaid with a suitable embedding medium (e. g. Mowiol, Hoechst) and examined under the fluorescence microscope. lf a POD-conjugate is used as the secondary antibody, the preparation should be overlaid with a substrate solution (see below) and incubated at 15-25°C until a clearly visible redbrown color develops. A negative control (e.g. only the labeled secondary antibody) should remain unchanged in color during this incubation period.

Subsequently, the substrate is washed off with PBS and the preparation is stained with hemalum stain for about 1 min. The hemalum solution is washed off with PBS, the preparation is embedded and examined.

Substrate solutions:

Aminoethyl-carbazole: Dissolve 2 mg 3-amino-9-ethylcarbazole with 1.2 mL dimethylsulfoxide and add 28.8 mL Tris-HCI, 0.05 M; pH 7.3; and 20 μL 3% H 2 O 2 , (w/v). Prepare solution freshly each day. Diaminobenzidine: Dissolve 25 mg 3, 3'-diaminobenzidine with 50 mL Tris-HCI, 0.05 M; pH 7.3; and add 40 μL 3% H2O2 , (w/v). Prepare solution freshly each day. Note The antibody solutions have to be absolutely free of precipitate! lf necessary, centrifuge the solutions at high speed prior to use.

Biological Information
Immunogen	Lung tumor cell lines A549 and A2182.
Clone	Lu5
Concentration	Please refer to the Certificate of Analysis for the lot-specific concentration.
Host	Mouse
Specificity	The antibody reacts with an epitope which is common to all cytokeratins respectively with a cytokeratin-associated protein. The antigen recognized by anti-cytokeratin pan is present in all types of epithelia. The antibody reacts with epithelial and mesothelial cells. Exception: Secretory cells are not stained and acinus cells, for example, show only weak staining. The antibody does not react with connective tissue, blood or lymphatic vessels, smooth or skeletal muscles. Exception: When higher antibody concentrations are used, weakly positive reactions can be observed with the smooth muscles of the myometrium, the prostata as well as the gastric muscularis mucosa.The antibodies can be used for differential diagnosis of epithelial tumors, distinction from mesenchymal, neural tumors as well as large-cell lymphomas. Anti-cytokeratin pan stains all epithelial tumors (primary tumors as well as metastases). Exception: Only partially positive staining is observed with theca cell tumors, granulosa cell tumors, adrenal cortex tumors. In mixed tumors (e.g. carcinosarkoma, thymoma, adenolymphoma) only the epithelial regions are stained. Tumors of nonepithelial origin do not react with the antibody. Exception: With high antibody concentrations a weak staining of leiomyosarcoma of the uterus is observed.
Isotype	IgG1
Species Reactivity	Mammals
Antibody Type	Monoclonal Antibody

Biological Information

Immunogen

Lung tumor cell lines A549 and A2182.

Clone

Lu5

Concentration

Please refer to the Certificate of Analysis for the lot-specific concentration.

Host

Mouse

Specificity

The antibody reacts with an epitope which is common to all cytokeratins respectively with a cytokeratin-associated protein. The antigen recognized by anti-cytokeratin pan is present in all types of epithelia. The antibody reacts with epithelial and mesothelial cells. Exception: Secretory cells are not stained and acinus cells, for example, show only weak staining. The antibody does not react with connective tissue, blood or lymphatic vessels, smooth or skeletal muscles. Exception: When higher antibody concentrations are used, weakly positive reactions can be observed with the smooth muscles of the myometrium, the prostata as well as the gastric muscularis mucosa.The antibodies can be used for differential diagnosis of epithelial tumors, distinction from mesenchymal, neural tumors as well as large-cell lymphomas. Anti-cytokeratin pan stains all epithelial tumors (primary tumors as well as metastases). Exception: Only partially positive staining is observed with theca cell tumors, granulosa cell tumors, adrenal cortex tumors. In mixed tumors (e.g. carcinosarkoma, thymoma, adenolymphoma) only the epithelial regions are stained. Tumors of nonepithelial origin do not react with the antibody. Exception: With high antibody concentrations a weak staining of leiomyosarcoma of the uterus is observed.

Isotype

IgG1

Species Reactivity

Mammals

Antibody Type

Monoclonal Antibody

Product Usage Statements
Usage Statement	Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Product Usage Statements

Usage Statement

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Storage and Shipping Information
Storage Conditions	The lyophilized antibody is stabe when stored at 2-8°C.The reconstituted antibody solution is stable at 2-8°C for three monthes. The solution can be stored in aliquots at -15 to -25 °C for 12 monthes. Avoid repeat freeze/thaw cycles.

Storage and Shipping Information

Storage Conditions

The lyophilized antibody is stabe when stored at 2-8°C.The reconstituted antibody solution is stable at 2-8°C for three monthes. The solution can be stored in aliquots at -15 to -25 °C for 12 monthes. Avoid repeat freeze/thaw cycles.

Packaging Information
Material Size	40 µg

Packaging Information

Material Size

40 µg

实验耗材

分子生物学试剂

蛋白与免疫学

生化试剂

细胞及检测试剂

色谱及层析

仪器设备

实验服务

办公用品

Anti-Cytokeratin Pan (mouse monoclonal)

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