Anti-Neurofilament light polypeptide (NF-L), DG-Sensor™ Antibody

Name: Anti-Neurofilament light polypeptide (NF-L), DG-Sensor™ Antibody
Brand: Biosensis
SKU: C-2125-100
Price: 327.0 USD
Availability: InStock

Our Anti-Neurofilament light polypeptide (NF-L) [degenerated] chicken polyclonal primary antibody detects bovine, human, mouse, pig, and rat Neurofilament light polypeptide (NF-L) [degenerated], and is IgG. It is validated for use in IF, IHC-non-Paraffin-embedded, WB.

Image of a rat spinal cord section with a three-day-old midline C4 contusion injury by Immunofluorescence and Immunohistochemistry. The section was stained with C-2125-100, Neurofilament light polypeptide (NF-L), DG-Sensor™, Chicken pAb, (green), and co-stained with product R-2113-50, Neurofilament light polypeptide, C-terminus, (NF-L-Ct), Rabbit pAb, (red). Product C-2125-100 stains prominent aggregates of material concentrated in the lateral funiculi and dorsal columns, which are degenerating and degenerated axons caused by the C4 lesion. As product R-2113-50 antibody binds the C-terminal "tail" region of NF-L, which is absent or destroyed during degeneration, the DG-Sensor™ C-2125 stains positively for degenerating NF-L profiles, but staining with R-2113-50 full-length NF-L is largely negative.

Analysis by western blot of NF-L expression in different tissue lysates with C-2125-100, Neurofilament light polypeptide (NF-L), DG-Sensor™, Chicken pAb, (green, 1:10,000) with strong bands at approx 68kDa. Lane 1: protein standard (red); Lane 2: rat brain; Lane 3: rat spinal cord; Lane 4: mouse brain; Lane 5: mouse spinal cord; Lane 6: bovine spinal cord; Lane 7: pig spinal cord. DG-Sensor™ antibodies detect NF-L in denatured protein lysates regardless of degeneration state. The lower molecular weight bands detected in the spinal cord samples are proteolytic artefacts

Image of a rat spinal cord section with a three-day-old midline C4 contusion injury by Immunofluorescence and Immunohistochemistry. The section was stained with C-2125-100, Neurofilament light polypeptide (NF-L), DG-Sensor™, Chicken pAb, (green), and co-stained with product R-2113-50, Neurofilament light polypeptide, C-terminus, (NF-L-Ct), Rabbit pAb, (red). Product C-2125-100 stains prominent aggregates of material concentrated in the lateral funiculi and dorsal columns, which are degenerating and degenerated axons caused by the C4 lesion. As product R-2113-50 antibody binds the C-terminal "tail" region of NF-L, which is absent or destroyed during degeneration, the DG-Sensor™ C-2125 stains positively for degenerating NF-L profiles, but staining with R-2113-50 full-length NF-L is largely negative.

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Species Reactivity

Bovine, Human, Mouse, Pig, Rat

Applications

IF, IHC, WB

Host Species

Chicken

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Data Sheet

SKU: C-2125-100

Volume/Size: 100 µg

Variant

Price:

$327.00

Quantity:

Product Details

Target

Neurofilament light polypeptide (NF-L) [degenerated]

Scientific Background

Our DG-Sensor™ chicken antibody works well on western blots of a variety of species for NF-L protein detection, regardless of injury state, but binds only degenerated processes in fixed, sectioned native material on injured or diseased brains. Caution: epitope reactivity is sensitive to protease attack; proteinase antigen recovery is not recommended and could lead to false positives in staining.

Format

IgG

Clonality

Polyclonal

Isotype

IgY

Applications

IF, IHC, WB

Host Species

Chicken

Immunogen

The antibody has been made against a proprietary recombinant construct containing amino acids of human NF-L expressed in and purified from E Coli.

Immunogen Species

Human

Species Reactivity

Bovine, Human, Mouse, Pig, Rat

Storage

Spin vial briefly before opening. Reconstitute with 100 µL sterile-filtered, ultrapure water to achieve a 1 mg/mL concentration. Centrifuge to remove any insoluble material. Store lyophilized antibody at 2-8°C After reconstitution of lyophilized antibody, aliquot and store at -20°C for a higher stability. Avoid freeze-thaw cycles. Store at 4°C for up to one month for short term storage and frequent use.

Physical State

Lyophilized

Production Notes

Affinity-purified

Buffer

Lyophilized from PBS buffer pH 7.2-7.6 with 0.1% trehalose, and sodium azide

Dilution Ranges

WB: 1:1000-1:5000
IHC: 1:1000-1:10000
ICC: 1:1000-1:10000

Application Details

CPCA-NF-L-Degen was raised against a proprietary recombinant immunogen containing amino acids 311-375 of the human NF-L sequence. The antibody works well on western blots of a variety of species but binds only degenerated processes in sectioned material.

Western blot (WB):

The antibody recognizes NF-L in reduced westerns regardless of the disease state. Caution: epitope reactivity is sensitive to protease attack; proteinase antigen recovery is not recommended and could lead to false positives in staining.

Immunofluorescence (IF), Immunohistochemistry (IHC):

Block and permeabilize sections in 5-10% normal goat serum or serum of the species the secondary antibodies were made in, in PBS plus 1% Triton-100 (PBST) for 1 hour with slight agitation, followed by primary antibody incubations and fluorescent secondary identification. High primary antibody dilutions require refrigerated, overnight incubations for best results. Recommended fixation is 4% PFA fixed, frozen tissue 20-50 microns; other fixation methods have not been tested and are not recommended at this time.

Degeneration-specific detection is fixation, antigen recovery, and concentration-dependent. C-2125- 100 was raised against a proprietary recombinant immunogen within the human NF-L sequence's amino acids 311-375. This peptide epitope can be uncovered in degenerating cells but not normal cells. However, treatment of the fixed tissue with high temperatures, proteinase, or other denaturants may cause the reactive epitope to become exposed in healthy cells, leading to a false positive. Biosensis recommends experimenting with treated and untreated tissues when first using these antibodies if degeneration specificity is desired. The exact conditions and dilutions must be determined experimentally by the end user.

This antibody will detect NL-L protein in paraffin-embedded rodent tissues; however, the degeneration-specific detection can be problematic in paraffin tissues, particularly if Heat-Induced Epitope Retrieval (HIER), or other common antigen recovery methods are used. This is because the reactive epitope, which is covered in healthy cells but exposed in degenerative cells, could become accessible to the antibody in healthy cells, leading to false positives. For this reason, paraffin- embedded tissues are not recommended if degeneration-specific detection is desired.

Conjugate

Unconjugated

Specificity

Species cross-reactivity includes human, rat, mouse, cow, and pig. These are antibodies which recognize epitopes in a small segment of the neurofilament NF-L subunit which are normally not accessible to antibodies but which became available on degeneration in natural or native conditions. Under reduced or proteinase treated tissues or Western blot, these antibodies are all specific for NF-L from a variety of species.

Usage Statement

For research use only.

Country of Origin

United States

Expiration

12 months after date of receipt (unopened vial).

Synonyms

NFL, Neurofilament light polypeptide, DG-Sensor, NF-L degenerative, NEFL, CMT1F, CMT2E, NF-L, NF68, PPP1R110, neurofilament light, CMTDIG, neurofilament light chain

UniProt ID

P07196

Shipping

25°C (ambient)

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Anti-Neurofilament light polypeptide (NF-L), DG-Sensor™ Antibody

SKU: C-2125-100

Product Details

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