Revision 6

#67255

Store at -20C

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, 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 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.

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).

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

W: 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

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

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
Western blot analysis of extracts from various tissues and cell lines using Neurofilament-M (E7L2T) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 1: Neurofilament-M (E7L2T) Rabbit mAb
Simple Western™ analysis of lysates (0.5 mg/mL) from Mouse Brain Tissue Extracts using Neurofilament-M (E7L2T) Rabbit mAb #67255. The virtual lane view (left) shows a single target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 66-440 kDa separation module.
Western Blotting Image 1: Neurofilament-M (E7L2T) Rabbit mAb
Immunoprecipitation of Neurofilament-M protein from mouse brain tissue extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Neurofilament-M (E7L2T) Rabbit mAb. Western blot analysis was performed using Neurofilament-M (E7L2T) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody.
Immunoprecipitation Image 1: Neurofilament-M (E7L2T) Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
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
Immunohistochemical analysis of paraffin-embedded normal human brain using Neurofilament-M (E7L2T) Rabbit mAb.
Immunohistochemistry Image 1: Neurofilament-M (E7L2T) Rabbit mAb
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Neurofilament-M (E7L2T) Rabbit mAb.
Immunohistochemistry Image 2: Neurofilament-M (E7L2T) Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using Neurofilament-M (E7L2T) Rabbit mAb.
Immunohistochemistry Image 3: Neurofilament-M (E7L2T) Rabbit mAb
Immunohistochemical analysis of paraffin-embedded normal human brain using Neurofilament-M (E7L2T) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Immunohistochemistry Image 4: Neurofilament-M (E7L2T) Rabbit mAb
Immunohistochemical analysis of paraffin-embedded normal mouse brain using Neurofilament-M (E7L2T) Rabbit mAb.
Immunohistochemistry Image 5: Neurofilament-M (E7L2T) Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded normal mouse stomach using Neurofilament-M (E7L2T) Rabbit mAb.
Immunohistochemistry Image 6: Neurofilament-M (E7L2T) Rabbit mAb
Immunohistochemical analysis of paraffin-embedded normal mouse thymus with adjacent nerve using Neurofilament-M (E7L2T) Rabbit mAb.
Immunohistochemistry Image 7: Neurofilament-M (E7L2T) Rabbit mAb
Immunohistochemical analysis of paraffin-embedded A-673 cell pellet (left, positive) or T24 cell pellet (right, negative) using Neurofilament-M (E7L2T) Rabbit mAb.
Immunohistochemistry Image 8: Neurofilament-M (E7L2T) Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
Confocal immunofluorescent analysis of mouse striatum using Neurofilament-M (E7L2T) Rabbit mAb (green), β3-Tubulin (TU-20) Mouse mAb #4466 (red), and ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 1: Neurofilament-M (E7L2T) Rabbit mAb
Confocal immunofluorescent analysis of mouse cerebellum using Neurofilament-M (E7L2T) Rabbit mAb (green), β3-Tubulin (TU-20) Mouse mAb #4466 (red), and ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 2: Neurofilament-M (E7L2T) Rabbit mAb
Confocal immunofluorescent analysis of A-673 cells (left, positive) or T24 cells (right, negative) using Neurofilament-M (E7L2T) Rabbit mAb (green), DyLight™ 650 Phalloidin #12956 (red), and DAPI #4083 (blue).
Immunofluorescence Image 1: Neurofilament-M (E7L2T) Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.