Revision 6
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, IP, IHC-Bond, IHC-P, IF-F, IF-IC

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

160

Source/Isotype:

Rabbit IgG

UniProt ID:

#P07197

Entrez-Gene Id:

4741

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunoprecipitation 1:50
IHC Leica Bond 1:200 - 1:800
Immunohistochemistry (Paraffin) 1:50 - 1:200
Immunofluorescence (Frozen) 1:200 - 1:800
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 #53682.

Specificity / Sensitivity

Neurofilament-M (E7L2T) Rabbit mAb recognizes endogenous levels of total Neurofilament-M protein. Non-specific staining was observed in skeletal muscle and some immune cells by immunohistochemistry.

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 BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation 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.
SignalStain is a registered 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 6
#67255

Neurofilament-M (E7L2T) Rabbit mAb

Western Blotting Image 1: Neurofilament-M (E7L2T) Rabbit mAb Expand Image
使用 Neurofilament-M (E7L2T) Rabbit mAb(上图)和 β-Actin (D6A8) Rabbit mAb #8457(下图)对不同组织和细胞系的提取物进行蛋白质印迹分析。
Western Blotting Image 1: Neurofilament-M (E7L2T) Rabbit mAb Expand Image
Simple Western™ analysis of lysates (0.5 mg/mL) from Mouse Brain Tissue Extracts using Neurofilament-M (E7L2T) Rabbit mAb #67255. 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 66-440 kDa 分离模块在 ProteinSimple(BioTechne 品牌)的 Jess™ Simple Western 仪器上进行该实验。
Immunoprecipitation Image 1: Neurofilament-M (E7L2T) Rabbit mAb Expand Image
对小鼠脑组织提取物的 Neurofilament-M 蛋白进行免疫沉淀分析。泳道 1 为 10% input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 Neurofilament-M (E7L2T) Rabbit mAb。使用 Neurofilament-M (E7L2T) Rabbit mAb 进行蛋白质印迹分析。Anti-rabbit IgG, HRP-linked Antibody #7074 用作二抗。
Immunohistochemistry Image 1: Neurofilament-M (E7L2T) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal human brain using Neurofilament-M (E7L2T) Rabbit mAb performed on the Leica BOND RX.
Immunohistochemistry Image 1: Neurofilament-M (E7L2T) Rabbit mAb Expand Image
使用 Neurofilament-M (E7L2T) Rabbit mAb 对石蜡包埋的正常人脑进行免疫组织化学分析。
Immunohistochemistry Image 2: Neurofilament-M (E7L2T) Rabbit mAb Expand Image
使用 Neurofilament-M (E7L2T) Rabbit mAb 对石蜡包埋的人结肠癌进行免疫组织化学分析。
Immunohistochemistry Image 3: Neurofilament-M (E7L2T) Rabbit mAb Expand Image
使用 Neurofilament-M (E7L2T) Rabbit mAb 对石蜡包埋的人前列腺癌进行免疫组织化学分析。
Immunohistochemistry Image 4: Neurofilament-M (E7L2T) Rabbit mAb Expand Image
以浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(右图)作为对照,使用 Neurofilament-M (E7L2T) Rabbit mAb(左图)对石蜡包埋的正常人脑进行免疫组织化学分析。
Immunohistochemistry Image 5: Neurofilament-M (E7L2T) Rabbit mAb Expand Image
使用 Neurofilament-M (E7L2T) Rabbit mAb 对石蜡包埋的正常小鼠脑进行免疫组织化学分析。
Immunohistochemistry Image 6: Neurofilament-M (E7L2T) Rabbit mAb Expand Image
使用 Neurofilament-M (E7L2T) Rabbit mAb 对石蜡包埋的正常小鼠胃进行免疫组织化学分析。
Immunohistochemistry Image 7: Neurofilament-M (E7L2T) Rabbit mAb Expand Image
使用 Neurofilament-M (E7L2T) Rabbit mAb 对石蜡包埋的正常小鼠胸腺及邻近神经进行免疫组织化学分析。
Immunohistochemistry Image 8: Neurofilament-M (E7L2T) Rabbit mAb Expand Image
使用 Neurofilament-M (E7L2T) Rabbit 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 (E7L2T) Rabbit mAb Expand Image
使用 Neurofilament-M (E7L2T) Rabbit mAb(绿色)、β3-Tubulin (TU-20) Mouse mAb #4466(红色)和 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)对小鼠纹状体进行共聚焦免疫荧光分析。
Immunofluorescence Image 2: Neurofilament-M (E7L2T) Rabbit mAb Expand Image
使用 Neurofilament-M (E7L2T) Rabbit mAb(绿色)、β3-Tubulin (TU-20) Mouse mAb #4466(红色)和 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)对小鼠小脑进行共聚焦免疫荧光分析。
Immunofluorescence Image 1: Neurofilament-M (E7L2T) Rabbit mAb Expand Image
使用 Neurofilament-M (E7L2T) Rabbit mAb(绿色)、DyLight 650 Phalloidin #12956(红色)和 DAPI #4083(蓝色)对 A-673 细胞(左图,阳性)或 T24 细胞(右图,阴性)进行共聚焦免疫荧光分析。