MAB3430
Anti-Desmin Antibody, clone DE-B-5
clone DE-B-5, Chemicon®, from mouse
About This Item
Produtos recomendados
fonte biológica
mouse
Nível de qualidade
forma do anticorpo
purified antibody
clone
DE-B-5, monoclonal
reatividade de espécies
mouse, frog, pig, rat, human
fabricante/nome comercial
Chemicon®
técnica(s)
immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable
western blot: suitable
Isotipo
IgG1
nº de adesão NCBI
nº de adesão UniProt
Condições de expedição
wet ice
modificação pós-traducional do alvo
unmodified
Informações sobre genes
human ... DES(1674)
Especificidade
Imunogênio
Aplicação
Optimal working dilutions must be determined by end user.
Immunohysto/cyto chemistry Protocols
Ideal specimens are obtained from frozen sections from shock-frozen tissue samples. The frozen sections are dried in the air and then fixed with acetone at -20°C for 10 min. Excess acetone is allowed to evaporate at 15-25°C. Material fixed in alcohol and embedded in paraffin can also be used (2). Formaldehyde fixation will reduce or eliminate the intensity of staining depending on the conditions under which it is performed. Other fixation conditions must be first tested by the investigator.
It 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 1 h.
• Dip the slide briefly in PBS and then wash 3 x 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 a solution of anti-mouse Ig-FITC or anti-mouse IgG-peroxidase solution 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.
If using an indirect immunofluorescence technique, the preparation should be overlaid with a suitable embedding medium (e.g. Moviol, Hoechst) and examined under the fluorescence microscope. If 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. Subsequently, the substrate is washed off with PBS and the preparation is stained, if desired, 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 50 mM Tris-HCI, 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 50 mM Tris-HCI, pH 7.3, and add 40 μl 3% H 2 O 2 (w/v). Prepare solution freshly each day.
Cell Structure
Cytoskeleton
Qualidade
Ligação
forma física
Armazenamento e estabilidade
Outras notas
Informações legais
Exoneração de responsabilidade
recomendado
Código de classe de armazenamento
12 - Non Combustible Liquids
Classe de risco de água (WGK)
WGK 2
Ponto de fulgor (°F)
Not applicable
Ponto de fulgor (°C)
Not applicable
Certificados de análise (COA)
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