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Horse BDNF(Brain Derived Neurotrophic Factor) ELISA Kit

SKU:
HRFI0007
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q0EAB7
Sensitivity:
18.75pg/ml
Range:
31.25-2000pg/ml
ELISA Type:
Sandwich
Synonyms:
BDNF, Brain Derived Neurotrophic Factor, ANON2, BULN2, Abrineurin, neurotrophin
Reactivity:
Horse
€649
Frequently bought together:

Description

Horse BDNF (Brain Derived Neurotrophic Factor) ELISA Kit

system_update_alt Datasheet system_update_alt MSDS

Key Features

Save Time Pre-coated 96 well plate
Quick Start Kit includes all necessary reagents
Publication Ready Reproducible and reliable results

Overview

Product Name:

Horse BDNF (Brain Derived Neurotrophic Factor) ELISA Kit

Product Code:

HRFI0007

Size:

96 Assays

Target:

Horse BDNF

Alias:

BDNF, Brain Derived Neurotrophic Factor, ANON2, BULN2, Abrineurin, neurotrophin

Application:

Horse

Detection Method:

Sandwich ELISA, Double Antibody

Sensitivity:

18.75pg/ml

Range:

31.25-2000pg/ml

Storage:

4°C for 6 months

Note:

For Research Use Only

Additional Information

Recovery:

Matrices listed below were spiked with certain level of Horse BDNF and the recovery rates were calculated by comparing the measured value to the expected amount of Horse BDNF in samples. Please contact us for more information.

Linearity:

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Horse BDNF and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected. Please get in contact for more information.

Intra-Assay:

CV <8%

Linearity:

CV <10%

Protein Information

Uniprot:

Q0EAB7

UniProt Protein Function:

During development, promotes the survival and differentiation of selected neuronal populations of the peripheral and central nervous systems. Participates in axonal growth, pathfinding and in the modulation of dendritic growth and morphology. Major regulator of synaptic transmission and plasticity at adult synapses in many regions of the CNS.

UniProt Code:

Q0EAB7

NCBI GenInfo Identifier:

122055974

NCBI Gene ID:

100009689

NCBI Accession:

Q0EAB7.1

UniProt Related Accession:

Q0EAB7

Molecular Weight:

27,722 Da

Protocol

*Note: Protocols are specific to each batch/lot. For the exact instructions please follow the protocol included in your kit.

Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37°C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.

Step Procedure

1.

Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!

2.

Aliquot 0.1ml standard solutions into the standard wells.

3.

Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.

4.

Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells.

5.

Seal the plate with a cover and incubate at 37 °C for 90 min.

6.

Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2.

7.

Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall.

8.

Seal the plate with a cover and incubate at 37°C for 60 min.

9.

Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash.

10.

Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min.

11.

Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.

12.

Add 90 µl of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color.

13.

Add 50 µl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.

14.

Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.

Sample Preparation

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample Type Protocol

Serum

If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

Plasma

Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.

Urine & Cerebrospinal Fluid

Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.

Cell culture supernatant

Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.

Cell lysates

Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.

Tissue homogenates

The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.

Tissue lysates

Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C

Breast Milk

Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

BDNF Background

BDNF, or Brain-Derived Neurotrophic Factor, is a protein that plays a crucial role in the growth, development, and maintenance of neurons in the brain and peripheral nervous system. BDNF is a member of the neurotrophin family of proteins.

Functions and Role of BDNF

BDNF is involved in various processes related to neuronal health and function. It promotes the survival of existing neurons, encourages the growth and differentiation of new neurons and synapses (connections between neurons), and enhances synaptic plasticity, which is the ability of the brain to change and adapt in response to experience.

BDNF in Neuronal Health and Development

Studies have shown that BDNF is essential for proper brain development during early life and continues to play a role in neuroplasticity throughout adulthood. It is involved in learning and memory processes and has been linked to various neurological and psychiatric disorders.

BDNF in Synaptic Plasticity and Adaptation

BDNF plays a critical role in synaptic plasticity, which is crucial for learning and memory formation. It influences the strength and structure of synapses, facilitating the communication between neurons and allowing for adaptive changes in neural circuits.

BDNF in Learning and Memory Processes

BDNF is closely associated with learning and memory processes. It promotes the growth and branching of dendrites, the receiving end of neurons, which enhances the formation and stability of synaptic connections. Additionally, BDNF is involved in the modulation of long-term potentiation (LTP), a cellular mechanism underlying memory formation.

BDNF in Neurological and Psychiatric Disorders

Imbalances or deficiencies in BDNF levels have been associated with conditions such as Alzheimer's disease, depression, anxiety disorders, schizophrenia, and neurodegenerative disorders. Conversely, increased BDNF levels have been observed in response to exercise, environmental enrichment, and certain medications, suggesting that BDNF may be involved in the beneficial effects of these interventions on brain health.

BDNF FAQs

What is the Horse BDNF ELISA Kit?

The Horse BDNF ELISA Kit is a specialized research tool that accurately measures Brain Derived Neurotrophic Factor levels in biological samples.

What are the benefits of using the Horse BDNF ELISA Kit?

Using the Horse BDNF ELISA Kit offers precise and reliable measurement of BDNF levels in horses. It provides a streamlined workflow and enables researchers to gain insights into equine brain health and cognitive function.

Where can I find more information about the Horse BDNF ELISA Kit?

For more detailed information about the Horse BDNF ELISA Kit, including technical specifications, performance characteristics, and ordering details, please refer to the product brochure or contact our customer support team. We are here to assist you with any inquiries you may have.

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