Revision 5

#53086Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, W-S, IHC-Bond, IHC-P, IF-F, IF-IC

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

160

Source/Isotype:

Mouse IgG2a kappa

UniProt ID:

#P07197

Entrez-Gene Id:

4741

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
IHC Leica Bond 1:200 - 1:800
Immunohistochemistry (Paraffin) 1:200 - 1:800
Immunofluorescence (Frozen) 1:100 - 1:400
Immunofluorescence (Immunocytochemistry) 1:200 - 1:800

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #60044.

Specificity / Sensitivity

Neurofilament-M (E7Y8W) Mouse mAb recognizes endogenous levels of total Neurofilament-M protein.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human Neurofilament-M protein.

Background

The cytoskeleton consists of three types of cytosolic fibers: actin microfilaments, intermediate filaments, and microtubules. Neurofilaments are the major intermediate filaments found in neurons and consist of light (NFL), medium (NFM), and heavy (NFH) subunits (1). Similar in structure to other intermediate filament proteins, neurofilaments have a globular amino-terminal head, a central α-helical rod domain, and a carboxy-terminal tail. A heterotetrameric unit (NFL-NFM and NFL-NFH) forms a protofilament, with eight protofilaments comprising the typical 10 nm intermediate filament (2). While neurofilaments are critical for radial axon growth and determine axon caliber, microtubules are involved in axon elongation. PKA phosphorylates the head domain of NFL and NFM to inhibit neurofilament assembly (3,4). Research studies have shown neurofilament accumulations in many human neurological disorders, including Parkinson's disease (in Lewy bodies along with α-synuclein), Alzheimer's disease, Charcot-Marie-Tooth disease, and Amyotrophic Lateral Sclerosis (ALS) (1).

  1. Al-Chalabi, A. and Miller, C.C. (2003) Bioessays 25, 346-55.
  2. Cohlberg, J.A. et al. (1995) J Biol Chem 270, 9334-9.
  3. Hisanaga, S. et al. (1994) Mol Biol Cell 5, 161-72.
  4. Sihag, R.K. et al. (1999) J Neurochem 72, 491-9.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting W-S: Simple Western™ IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 5
#53086

Neurofilament-M (E7Y8W) Mouse mAb

Western Blotting Image 1: Neurofilament-M (E7Y8W) Mouse mAb Expand Image
使用 Neurofilament-M (E7Y8W) Mouse mAb(上图)或 α-Actinin (D6F6) XP® Rabbit mAb #6487(下图)对来自各种组织的提取物进行蛋白质印迹分析。神经丝-M 蛋白在小鼠胰腺组织中阴性表达与预测的表达模式一致。
Western Blotting Image 1: Neurofilament-M (E7Y8W) Mouse mAb Expand Image
Simple Western™ analysis of lysates (0.5 mg/mL) from Mouse Brain Tissue Extracts using Neurofilament-M (E7Y8W) Mouse mAb #53086. 虚拟泳道式图像(左图)显示一抗稀释比例为 1:10 和 1:50 时的单一靶标条带(如图所示)。对应的电泳图(右图)为一抗稀释比例在 1:10(蓝线)和 1:50(绿线)时沿毛细血管内分子量的化学发光结果。在还原条件下,使用 12-230 kDa 分离模块在 ProteinSimple(BioTechne 品牌)的 Jess™ Simple Western 仪器上进行该实验。
Immunohistochemistry Image 1: Neurofilament-M (E7Y8W) Mouse mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal human brain using Neurofilament-M (E7Y8W) Mouse mAb performed on the Leica BOND RX.
Immunohistochemistry Image 1: Neurofilament-M (E7Y8W) Mouse mAb Expand Image

使用 Neurofilament-M (E7Y8W) Mouse mAb 对石蜡包埋的人正常基底神经节组织进行免疫组织化学分析。

Immunohistochemistry Image 2: Neurofilament-M (E7Y8W) Mouse mAb Expand Image

使用 Neurofilament-M (E7Y8W) Mouse mAb 对石蜡包埋的人正常尾状核组织进行免疫组织化学分析。

Immunohistochemistry Image 3: Neurofilament-M (E7Y8W) Mouse mAb Expand Image
使用 Neurofilament-M (E7Y8W) Mouse mAb 对石蜡包埋的人正常基底神经节组织进行免疫组织化学分析。
Immunohistochemistry Image 4: Neurofilament-M (E7Y8W) Mouse mAb Expand Image
使用 Neurofilament-M (E7Y8W) Mouse mAb 对人前列腺腺癌内石蜡包埋的周围神经组织进行免疫组织化学分析。
Immunohistochemistry Image 5: Neurofilament-M (E7Y8W) Mouse mAb Expand Image
使用 Neurofilament-M (E7Y8W) Mouse mAb 对石蜡包埋的人正常心脏组织进行免疫组织化学分析。注释 周围神经染色。
Immunohistochemistry Image 6: Neurofilament-M (E7Y8W) Mouse mAb Expand Image
与浓度匹配的 Mouse (E5Y6Q) mAb IgG2a Isotype Control #61656(右图)比较,使用 Neurofilament-M (E7Y8W) Mouse mAb(左图)对石蜡包埋的人正常脑组织进行免疫组织化学分析。
Immunohistochemistry Image 7: Neurofilament-M (E7Y8W) Mouse mAb Expand Image
使用 Neurofilament-M (E7Y8W) Mouse mAb 对石蜡包埋的 A-673 细胞沉淀物(左图,阳性)或 T24 细胞沉淀物(右图,阴性)进行免疫组织化学分析。
Immunofluorescence Image 1: Neurofilament-M (E7Y8W) Mouse mAb Expand Image
对使用 Neurofilament-M (E7Y8W) Mouse mAb(左图,绿色)标记并使用 Iba1/AIF-1 (E4O4W) XP® Rabbit mAb (Alexa Fluor® 555 Conjugate) #36618(右图,伪彩青色)、GFAP (GA5) Mouse mAb (Alexa Fluor® 647 Conjugate) #3657(右图,红色)和 ProLong Gold Antifade Reagent with DAPI #8961(右图,蓝色)共同标记的固定冷冻的小鼠海马组织进行共聚焦免疫荧光分析。
Immunofluorescence Image 1: Neurofilament-M (E7Y8W) Mouse mAb Expand Image
使用 Neurofilament-M (E7Y8W) Mouse mAb(绿色)、DyLight 650 Phalloidin #12956(红色)和 DAPI #4083(蓝色)对 A-673 细胞(左图,阳性)和 T24 细胞(右图,阴性)进行共聚焦免疫荧光分析。