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

Name: Anti-Neurofilament light polypeptide (NF-L), DG-Sensor™ Antibody
Brand: Biosensis
SKU: R-2124-100
Price: 327.0 USD
Availability: InStock

Our Anti-Neurofilament light polypeptide (NF-L) [degenerated] rabbit 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 ELISA, IF, ICC, IHC, 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 R-2124-100, Neurofilament light polypeptide (NF-L) DG-Sensor™, Rabbit pAb, (red, 1:1,000) and co-stained with product M-1394-100, Neurofilament medium polypeptide (NF-M), Clone 3H11, Mouse mAb, (green, 1:1000). Product R-2124-100 does not stain the undamaged axons that M-1394-100 strongly stains (green). Linear arrays of swollen profiles originating from damaged axons, on the other hand, are strongly positive for R-2124-100, but not for the M-1394-100. In these regions, the M-1394-100 epitope, which is located in the C-terminal "tail" of NF-M, has been either removed or destroyed.

Analysis by western blot of NF-L expression in different tissue lysates with R-2124-100, Neurofilament light polypeptide (NF-L), DG-Sensor™, Rabbit pAb, (green, 1:2,000) with a band at approx 68-70kDa. 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.

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 R-2124-100, Neurofilament light polypeptide (NF-L) DG-Sensor™, Rabbit pAb, (red, 1:1,000) and co-stained with product M-1394-100, Neurofilament medium polypeptide (NF-M), Clone 3H11, Mouse mAb, (green, 1:1000). Product R-2124-100 does not stain the undamaged axons that M-1394-100 strongly stains (green). Linear arrays of swollen profiles originating from damaged axons, on the other hand, are strongly positive for R-2124-100, but not for the M-1394-100. In these regions, the M-1394-100 epitope, which is located in the C-terminal "tail" of NF-M, has been either removed or destroyed.

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Species Reactivity

Bovine, Human, Mouse, Pig, Rat

Applications

ELISA, ICC, IF, IHC, WB

Host Species

Rabbit

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Data Sheet

SKU: R-2124-100

Volume/Size: 100 µg

Variant

Price:

$327.00

Quantity:

Product Details

Target

Neurofilament light polypeptide (NF-L) [degenerated]

Scientific Background

Our DG-Sensor™ rabbit 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 sectioned 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

IgG

Applications

ELISA, ICC, IF, IHC, WB

Host Species

Rabbit

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. 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: 1:1000-1:2000
ICC: 1:1000

Application Details

The antibody recognizes NF-L in reduced westerns regardless of the disease state.

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 and antigen recovery dependent. This antibody reacts to a peptide epitope within amino acids 311-375 of the human NF-L. This peptide epitope is uncovered in degenerating cells but not normal cells. However, the specific reactivity of this epitope is sensitive to tissue treatments and could become exposed in healthy cells under some conditions. For example, 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 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: R-2124-100

Product Details

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