MAB3430
Anti-Desmin Antibody, clone DE-B-5
clone DE-B-5, Chemicon®, from mouse
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About This Item
Código UNSPSC:
12352203
eCl@ss:
32160702
NACRES:
NA.41
Productos recomendados
origen biológico
mouse
Nivel de calidad
forma del anticuerpo
purified antibody
clon
DE-B-5, monoclonal
reactividad de especies
mouse, frog, pig, rat, human
fabricante / nombre comercial
Chemicon®
técnicas
immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable
western blot: suitable
isotipo
IgG1
Nº de acceso NCBI
Nº de acceso UniProt
Condiciones de envío
wet ice
modificación del objetivo postraduccional
unmodified
Información sobre el gen
human ... DES(1674)
Especificidad
The antibody reacts with desmin from human, pig, rat and taod. In tissue sections this antibody is used to stain skeletal, cardiac, visceral, and some vascular smooth muscle cells. Cell lines such as RD (ATCC CCL 136) and hamster BHK-21 are positive (Debus et al., 1983).
Inmunógeno
Purified desmin.
Aplicación
Detect Desmin using this Anti-Desmin Antibody, clone DE-B-5 validated for use in WB, IH, IH(P), IH(P).
Immunocytochemistry: (5 μg/ml) Immunohistochemistry: (5 μg/ml) See IHC2011-6 for prediluted and detailed paraffin protocols.
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.
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.
Research Category
Cell Structure
Cell Structure
Research Sub Category
Cytoskeleton
Cytoskeleton
Calidad
Western Blot Analysis: 1:1000 dilution
Ligadura / enlace
Replaces: 04-585
Forma física
Format: Purified
Liquid in 0.02 M phosphate buffer, 0.25M NaCl with 0.1% sodium azide, pH 7.6.
Almacenamiento y estabilidad
Maintain antibody refrigerated at 2-8°C in undiluted aliquots for up to 6 months. DO NOT FREEZE.
Otras notas
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Información legal
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Cláusula de descargo de responsabilidad
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.
Opcional
Referencia del producto
Descripción
Precios
Código de clase de almacenamiento
12 - Non Combustible Liquids
Clase de riesgo para el agua (WGK)
WGK 2
Punto de inflamabilidad (°F)
Not applicable
Punto de inflamabilidad (°C)
Not applicable
Certificados de análisis (COA)
Busque Certificados de análisis (COA) introduciendo el número de lote del producto. Los números de lote se encuentran en la etiqueta del producto después de las palabras «Lot» o «Batch»
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