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Anti-Neurofilament 200 kDa Antibody, clone N52

产品编号:4110882
规格:clone N52, Chemicon®, from mouse
包装规格:50 μG
产品类别:进口试剂
品牌:Sigma-Aldrich
优惠价:立即咨询
产品价格
产品编号包装单位单价(元)国内现货国外库存询价单
411088250 μG5520
基本信息
eCl@ss
32160702
NACRES
NA.41
General description【一般描述】
Neurofilaments are a type of intermediate filament that serve as major elements of the cytoskeleton supporting the axon cytoplasm. They are the most abundant fibrillar components of the axon, being on average 3-10 times more frequent than axonal microtubules. Neurofilaments (10nm in dia.) are built from three intertwined protofibrils which are themselves composed of two tetrameric protofilament complexs of monomeric proteins. The neurofilament triplet proteins (68/70, 160, and 200 kDa) occur in both the central and peripheral nervous system and are usually neuron specific. The 68/70 kDa NF-L protein can self-assemble into a filamentous structure, however the 160 kDa NF-M and 200 kDa NF-H proteins require the presence of the 68/70 kDa NF-L protein to co-assemble. Neuromas, ganglioneuromas, gangliogliomas, ganglioneuroblastomas and neuroblastomas stain positively for neurofilaments. Although typically restricted to neurons, neurofilaments have been detected in paragangliomas and adrenal and extra-adrenal pheochromocytomas. Carcinoids, neuroendocrine carcinomas of the skin, and oat cell carcinomas of the lung also express neurofilaments. For more neurofilament information see Nervous System Cell Type Specific Marker chart online under the CHEMICON Technical Support section.
Specificity【特异性】
MAB5266 reacts with the phophorylated and dephosphorylated H-chain of neurofilament 200kDa (NF-H) in normal tissues/extracts (Shaw, 1986). MAB5266 can be used to detect cells of neuronal origin by immunohistochemistry and Western blot. It has been reported that reactivity of MAB5266 with NF-H is blocked in cdk-5 over-expressing cells (Guidato, 1996).
Immunogen【免疫原】
Carboxy terminal tail segment of enzymatically dephosphorylated porcine H-chain.
Application【应用】
Research Category
Neuroscience
Research Sub Category
Neurofilament & Neuron Metabolism

Neuronal & Glial Markers
Western blot: 5-10 μg/mL

Immunohistochemistry: 5-10 μg/mL

Optimal working dilutions must be determined by end user.

Immunohistochemistry: Antibody N52 reacts with a fragment of NF-200 side-arm that is located at the end of the MPR KSP domain and which contains the consensus cdk-5 phosphorylation site, however this reactivity is abolished if the NF-200 fragment becomes phosphorylated by cdk-5 {Guidato et al, 1996}. Thus for the fullest staining and reactivity, including westerns, it is suggested that samples be treated with alkaline phosphatase prior to antibody staining. The following treatment protocol is suggested:



ProtocolPreparation of Solutions:

I. TBS: Tris-HCl, 0.1 mol/l; NaCl, 0.1 mol/l′ pH 8.0.

II. Mix alkaline phosphatase, 1 mg/ml with 1 mol/l ZnSO4,; 1mol/l MgCl2, and 1mmol/l PMSF; and dissove in TBS.

NOTE: It is necessary to dialyze the alkaline phosphatase against solution I before use.

Procedure:

Frozen sections from shock frozen tissue samples are air-dried and then fixed with acetone for 10 min at -20°C. The excess acetone is allowed to evaporate at room temperature. Incubate the sections in solutions II for 4 hours at +30oC and wash in TBS 3 times for 5 minutes each. Negative control sections are incubated in solution II with out alkaline phosphatase. Further treatment as follows:

- Cover the preparation with 10-20 μl antibody solution and incubate for 1 hr in a humid chamber.

- Immerse the slide in TBS and wash in TBS 3 times for 5 minutes each.

- Cover the preparation with 10-20 μl of a solution of anti-mouse Ig-antibody, labeled with fluorscine isothiocyanate, and incubate in a humid chamber at 37°C for 1 hour.

- Wash the slide in TBS 3 times for 5 min each.
This Anti-Neurofilament 200 kDa Antibody, clone N52 is validated for use in WB, IH for the detection of Neurofilament 200 kDa.
Physical form【外形】
Purified immunoglobulin (Ion exchange chromatography). Liquid in 0.02M phosphate buffer, 0.25M NaCl with 0.1% sodium azide, pH 7.6
Format: Purified
Other Notes【其他说明】
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Legal Information【法律信息】
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
产品性质
Quality Level【质量水平】
300
100
biological source【生物来源】
mouse
antibody form【抗体形式】
purified immunoglobulin
antibody product type
primary antibodies
clone【克隆】
N52, monoclonal
species reactivity
pig, mouse, human, monkey, rat
manufacturer/tradename
Chemicon®
technique(s)
immunohistochemistry: suitable
western blot: suitable
isotype【同位素/亚型】
IgG1
NCBI accession no.【NCBI登记号】
NM_021076.2
UniProt accession no.【UniProt登记号】
P12036
shipped in【运输】
wet ice
产品说明
Storage and Stability【储存及稳定性】
Maintain at 2-8°C in undiluted aliquots for up to 6 months. Avoid repeated freeze/thaw cycles.
Disclaimer【免责声明】
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.
安全信息
Storage Class Code【储存分类代码】
10 - Combustible liquids
WGK
WGK 2
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