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Key Documents

MAB3404

Sigma-Aldrich

Anti-Cytokeratin 18 Antibody, clone CK2

clone CK2, Chemicon®, from mouse

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About This Item

UNSPSC 코드:
12352203
eCl@ss:
32160702
NACRES:
NA.41

생물학적 소스

mouse

Quality Level

항체 형태

purified immunoglobulin

항체 생산 유형

primary antibodies

클론

CK2, monoclonal

종 반응성

human

제조업체/상표

Chemicon®

기술

immunocytochemistry: suitable
immunohistochemistry: suitable

동형

IgG1

NCBI 수납 번호

UniProt 수납 번호

배송 상태

wet ice

타겟 번역 후 변형

unmodified

유전자 정보

human ... KRT18(3875)

특이성

The antibody reacts with cytokeratin No. 18 from human. This antibody is used for staining research samples of a wide variety of simple epithelia and simple glandular epithelia (e. g. intestinal, respiratory and urinary epithelia) and transitional epithelia (e. g. bladder), but stratified squamous epithelia are not stained (e. g. esophagus, epidermis). Keratin filaments of cell lines such as HeLa cells are stained (Debus et al., 1982). For staining research samples of human tumor material with this antibody, see (Debus et al., 1984). For the cytokeratin pattern of different tissues, see (Moll et al., 1982).

면역원

Cytoskeletal preparation of HeLa cells (Debus et al., 1982).

애플리케이션

Immunocytochemistry: 20 μg/mL

Immunohistochemistry: 20 μg/mL Not for formaldehyde fixed tissues.

Optimal working dilutions must be determined by end user.

Immunohistochemistry/Immunocytochemistry Protocols:

Ideal specimens are obtained from frozen sections from shockfrozen tissue samples. The frozen sections are dried in the air and then fixed with acetone at -15 to -25°C for 10 min. Excess acetone is allowed to evaporate at 15-25°C. Material fixed in alcohol and embedded in paraffin can, however, be used, see (4). For the immunohistochemical detection of cytokeratin No. 18 in tissue sections, the tissue should not be fixed in formaldehyde as this fixative markedly reduces the staining intensity of the cytokeratin filaments. lt is advantageous to block unspecific binding sites by overlaying the sections with fetal calf serum for 20-30 min at 15-25°C. Excess of fetal calf serum is removed by decanting before application of the antibody solution.

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

• Overlay the preparation with 10-20 μL antibody solution and incubate in a humid chamber at 37°C for 1h.

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

• Wipe the margins of the preparation dry and overlay the preparation with 10-20 μL of an anti-mouse-IgG-FITC or anti-mouse-IgG-peroxidase antibody and allow to incubate for 1 h at 37°C in a humid chamber.

• Wash the slide 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 embed-ding medium (e. g. Moviol, Hoechst) and examined under the fluorescence microscope.

lf a POD-conjugate has been 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 secondary antibody) should remain unchanged in color during this incubation period.

Wash away the substrate with PBS and stain the preparation, if desired, with hemalum stain, for about 1 min. The hemalum solution is washed off with PBS, the peparation is embedded and examined.

Substrate solutions:

Aminoethyl-carbazole Dissolve 2mg 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 H 2 O 2 , 3% (w/v). Prepare solution freshly each day.
This Anti-Cytokeratin 18 Antibody, clone CK2 is validated for use in IC, IH for the detection of Cytokeratin 18.

결합

Replaces: 04-586

물리적 형태

Format: Purified

저장 및 안정성

The lyophilized antibody is stable when stored at -20°C.Store the reconstituted antibody solution at 2-8°C, for up to 12 months.DO NOT FREEZE.

기타 정보

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

법적 정보

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

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Storage Class Code

10 - Combustible liquids

WGK

WGK 2

Flash Point (°F)

Not applicable

Flash Point (°C)

Not applicable


시험 성적서(COA)

제품의 로트/배치 번호를 입력하여 시험 성적서(COA)을 검색하십시오. 로트 및 배치 번호는 제품 라벨에 있는 ‘로트’ 또는 ‘배치’라는 용어 뒤에서 찾을 수 있습니다.

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문서 라이브러리 방문

Characterization of liver cytokeratin as a major target antigen of anti-SLA antibodies.
Wachter, B, et al.
Journal of Hepatology, 11, 232-239 (1990)
[Monoclonal antibodies--new probes for diagnosis and therapy. Their use as an example of the micrometastasizing of solid tumors]
Schlimok, G, et al.
Arzneimittelforschung, 38, 435-437 (1988)
T S Weiss et al.
Gut, 57(8), 1129-1138 (2008-04-18)
Liver regeneration is mainly based on cellular self-renewal including progenitor cells. Efforts have been made to harness this potential for cell transplantation, but shortage of hepatocytes and premature differentiated progenitor cells from extra-hepatic organs are limiting factors. Histological studies implied
E Debus et al.
The American journal of pathology, 114(1), 121-130 (1984-01-01)
Carcinomas of different origin have been tested in immunofluorescence microscopy with the monoclonal murine antibodies CK1-CK4, which recognize a single cytokeratin polypeptide (human cytokeratin No. 18) present in simple but not in stratified squamous epithelia, and with the monoclonal antibody
E Debus et al.
The EMBO journal, 1(12), 1641-1647 (1982-01-01)
Four monoclonal antibodies designated CK1 - CK4 were obtained from fusions of mouse myeloma F0 cells with spleen cells from BALB/c mice immunized with cytoskeletal preparations made by treatment of human HeLa cells with non-ionic detergents. These IgG1 type antibodies

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