NF-M Polyclonal antibody

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  WB analysis using 25805-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 25805-1-AP (NF-M antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

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  WB analysis of mouse brain using 25805-1-AP

  mouse brain tissue were subjected to SDS PAGE followed by western blot with 25805-1-AP (NEFM Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  WB analysis of rat brain using 25805-1-AP

  rat brain tissue were subjected to SDS PAGE followed by western blot with 25805-1-AP (NEFM Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  IHC staining of mouse cerebellum using 25805-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue slide using 25805-1-AP (NF-M antibody) at dilution of 1:200 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse cerebellum using 25805-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue slide using 25805-1-AP (NF-M antibody) at dilution of 1:200 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human brain using 25805-1-AP

  Immunohistochemical analysis of paraffin-embedded human brain tissue slide using 25805-1-AP (NEFM Antibody) at dilution of 1:200 (under 10x lens).

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  IHC staining of human brain using 25805-1-AP

  Immunohistochemical analysis of paraffin-embedded human brain tissue slide using 25805-1-AP (NEFM Antibody) at dilution of 1:200 (under 40x lens).

• 

  IHC staining of mouse cerebellum using 25805-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue slide using 25805-1-AP (NF-M antibody) at dilution of 1:200 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse brain using 25805-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 25805-1-AP (NEFM Antibody) at dilution of 1:50 (under 40x lens).

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  IHC staining of human colon using 25805-1-AP

  Immunohistochemical analysis of paraffin-embedded human colon tissue slide using 25805-1-AP (NEFM Antibody) at dilution of 1:200 (under 10x lens).

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  IHC staining of human colon using 25805-1-AP

  Immunohistochemical analysis of paraffin-embedded human colon tissue slide using 25805-1-AP (NEFM Antibody) at dilution of 1:200 (under 40x lens).

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  IF Staining of mouse brain using 25805-1-AP

  Immunofluorescent analysis of (4% PFA) fixed mouse brain tissue using 25805-1-AP (NF-M antibody) at dilution of 1:50 and Alexa Fluor 488-Conjugated Goat Anti-Rabbit IgG(H+L).

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  FC experiment of PC-12 using 25805-1-AP

  1X10^6 PC-12 cells were intracellularly stained with 0.4 ug Anti-Human NF-M (25805-1-AP) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Control Antibody. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

NF-M Polyclonal Antibody for WB, IHC, IF-P, FC (Intra), ELISA

Cat No. 25805-1-AP

Publications(12)

Host / Isotype

Rabbit / IgG

Reactivity

human, rat, mouse and More (1)

Applications

WB, IHC, IF-P, FC (Intra), ELISA

Synonyms

NEFM, NF M, neurofilament, NEF3

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Tested Applications

• Positive WB detected in: mouse brain tissue, rat brain tissue
• Positive IHC detected in: mouse cerebellum tissue, human brain tissue, human colon tissue, mouse brain tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF-P detected in: mouse brain tissue
• Positive FC (Intra) detected in: PC-12 cells
• Positive FC detected in: PC-12 cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:10000
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF)-P: IF-P : 1:50-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension
• Flow Cytometry (FC): FC : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 7 publications below
• IHC: See 2 publications below
• IF: See 5 publications below

Product Information

25805-1-AP targets NF-M in WB, IHC, IF-P, FC (Intra), ELISA applications and shows reactivity with human, rat, mouse samples.

• Tested Reactivity: human, rat, mouse
• Cited Reactivity: human, mouse, rat, canine
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: NF-M fusion protein Ag22709
• Full Name: neurofilament, medium polypeptide
• Calculated Molecular Weight: 102 kDa
• Observed Molecular Weight: 140 kDa
• GenBank Accession Number: BC002421
• Gene Symbol: NF-M
• Gene ID (NCBI): 4741
• RRID: AB_2880245
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• UNIPROT ID: P07197
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

NEFM, also named as NEF3 and NFM, belongs to the intermediate filament family. Neurofilaments are the 10nm intermediate filaments found specifically in neurons. They are a major component of the cell's cytoskeleton, and provide support for normal axonal radial growth. Neurofilaments usually contain three intermediate filament proteins: L, M, and H which are involved in the maintenance of neuronal caliber. The names given to the three major neurofilament subunits are based upon the apparent molecular weight of the mammalian subunits on SDS-PAGE:NF-L, 65-68 kDa; NF-M,145-160 kDa and NF-H, 200-220 kDa. This antibody recognizes endogenous NF-M protein.

Publications

Species	Application	Title
canine	IF	Theranostics; Implantation of regenerative complexes in traumatic brain injury canine models enhances the reconstruction of neural networks and motor function recovery.; Authors - Jipeng Jiang
mouse, canine	IF	Biomater Sci; 3D printing collagen/heparin sulfate scaffolds boost neural network reconstruction and motor function recovery after traumatic brain injury in canine.; Authors - Jipeng Jiang
mouse	WB	Mol Autism; RNA sequencing and proteomics approaches reveal novel deficits in the cortex of Mecp2-deficient mice, a model for Rett syndrome.; Authors - Natasha L Pacheco
canine	IHC	Front Cardiovasc Med; Vagus Nerve Stimulation in Early Stage of Acute Myocardial Infarction Prevent Ventricular Arrhythmias and Cardiac Remodeling.; Authors - Shuang Zhao
rat	WB	Neurosci Lett; Overexpression of miR-338-5p in exosomes derived from mesenchymal stromal cells provides neuroprotective effects by the Cnr1/Rap1/Akt pathway after spinal cord injury in rats.; Authors - Anwei Zhang
rat	WB	Cell Biol Int; Study of the protective effect on damaged intestinal epithelial cells of rat multilineage-differentiating stress-enduring (Muse) cells.; Authors - Dong Sun