Rat Beta-NGF Rapid ELISA Assay

Name: Rat Beta-NGF Rapid ELISA Assay
Brand: Biosensis
SKU: BEK-2214-2P
Price: 787.0 USD
Availability: InStock

The Biosensis NGF Rapid enzyme-linked immunosorbent assay (ELISA) Kit is a sandwich ELISA that allows the quantification of rat NGF in less than 3 hours in cell culture supernatants, serum, and brain extracts. This NGF ELISA kit is designed to measure rat NGF and thus employs a recombinant rat NGF protein. Due to a high degree of NGF sequence homology, the antibodies used in this kit will also detect NGF from other species including human and mouse.

This standard curve generated in our laboratories is for demonstration purposes only, but can be used as a guide to expected performance. A standard curve should be generated for each assay.

BEK-2212 detects only NGF and cannot detect proNGF, as the capture antibody is specific for NGF and does not cross-react with the prodomain of the NGF.

This standard curve generated in our laboratories is for demonstration purposes only, but can be used as a guide to expected performance. A standard curve should be generated for each assay.

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Species Reactivity

Guinea Pig, Rat

Applications

ELISA

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Data Sheet

SKU: BEK-2214-2P

Volume/Size: 2 Plate Kit

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Product Description

The Biosensis NGF Rapid enzyme-linked immunosorbent assay (ELISA) Kit is a sandwich ELISA that allows the quantification of rat NGF in less than 3 hours in cell culture supernatants, serum, and brain extracts only if used as directed. Please refer to the kit protocol for specific use instructions for each substrate application, in particular serum samples.

This ELISA kit consists of a pre-coated mouse monoclonal anti-NGF capture antibody, a biotinylated anti-NGF detection antibody and horseradish peroxidase (HRP)-conjugated streptavidin. The addition of a substrate (3,3',5,5'-tetramethylbenzidine, TMB) yields a colored reaction product which is directly proportional to the concentration of NGF present in samples and protein standards.

This NGF ELISA kit is designed to measure rat NGF and thus employs a recombinant rat NGF protein. Due to a high degree of NGF sequence homology, the antibodies used in this kit will also detect NGF from other species including human and mouse! Guinea pig NGF has been quantified in serum using this ELISA kit using rat NGF protein as calibrator, as it shows largest sequence homology based on amino acid sequence among rodent and human NGF proteins.

The assay antibodies preferentially detect the mature form of NGF, as shown by data comparing mature mouse NGF and mouse proNGF in the Mouse NGF ELISA kit (BEK-2213). Cross-reaction of full-length proNGF was < 0.1% when assayed at 25 ng/mL in assay buffer, and not detectable when assayed across the mature NGF calibration range.

This kit has not been tested for other applications. It has been configured for research use only and is not to be used in diagnostic or clinical procedures.

Scientific Background

FUNCTION: Nerve growth factor is important for the development and maintenance of the sympathetic and sensory nervous systems. It stimulates division and differentiation of sympathetic and embryonic sensory neurons. SUBUNIT: Homodimer, associated by non-covalent forces. SUBCELLULAR LOCATION: Secreted protein. SIMILARITY: Belongs to the NGF-beta family.

Target

Nerve growth factor (Beta-NGF)

Immunogen

Sf 21 (baculovirus)-derived rat beta-NGF protein

Immunogen Species

Rat

Range

3.9 - 250 pg/mL

Sensitivity

Typical limit of detection (LOD) for rat NGF is 2 pg/mL determined as 150% of the blank value.

Specificity

The antibodies used in this ELISA kit bind epitopes within the mature domain of the protein. No cross-reactivity was observed with brain derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), NT-4/5 and proNGF protein tested at 25 ng/mL in assay buffer. Due to a high degree of NGF sequence homology, the antibodies used in this kit will also detect NGF from other species including human and mouse. Mature NGF (27 kDa) and full-length proNGF (50 kDa) were assayed in parallel at equimolar protein concentrations across the NGF ELISA calibration range (3.9-250 pg/mL; 0.14-9.2 pmol/L). OD readings for proNGF were indistinguishable from the assay's blank OD readings. Data was obtained with the Mouse NGF ELISA kit (BEK-2213) which uses the same assay capture and detection antibodies.

Species Reactivity

Guinea Pig, Rat

Applications

ELISA

Method of Analysis

Colorimetric

Sample Type

Culture Supernatant, Serum, Tissue Homogenates

Synonyms

Beta-nerve growth factor; Ngfb; NGF

UniProt ID

P25427

Usage Statement

For research use only.

Country of Origin

United States

Expiration

12 months from purchase.

Storage

Store at 2-8°C

Shipping

Shipped on ice packs

Rat Beta-NGF Rapid ELISA Assay Protocol
Technical Note 1 - ELISA Validation
Technical Note 2 - ELISA Data Analysis
Calculation of Antigen Amount in Tissues and Lysates

Reagent	1 Plate Kit	2 Plate Kit
NGF antibody coated 96 well microplate	12 × 8 wells	24 × 8 wells
Assay diluent A (1x)	2 × 25 mL	4 × 25 mL
Recombinant rat NGF standard	2 × 500 pg	4 × 500 pg
Quality Control (QC) Sample	2 Vials (see vial label for amount)	4 Vials (see vial label for amount)
NGF detection antibody (100x)	1 × 110 µL	2 × 110 µL
Streptavidin-HRP (100x)	1 × 110 µL	2 × 110 µL
Wash buffer (10x)	1 × 33 mL	2 × 33 mL
TMB substrate (1x)	1 × 11 mL	2 × 11 mL
TMB stop solution (1x)	1 × 11 mL	2 × 11 mL
Plate sealer	Supplied

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Rat Beta-NGF Rapid ELISA Assay

SKU: BEK-2214-2P

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