BDNF ELISA Kit (Rat) (OKBB00128)

Catalog No: OKBB00128 (Formerly GWB-ZZD023)

Size:96 Tests

Rat BDNF ELISA Kit standard curve

Product Info

• Predicted Species Reactivity: Rat
• Application: ELISA
• ELISA Kit Detection Method: Colorimetric, OD 450 nm
• ELISA Kit Duration: ~ 3 Hours
• ELISA Kit Principle: Aviva's rat BDNF ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for BDNF has been precoated onto 96-well plates. Standards(NS0, H129-R247) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for BDNF is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The absorbance of yellow is proportional to the rat BDNF amount of sample captured in plate.
• ELISA Kit Range: 31.2 pg/ml - 2,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (pg/mL)	109	447	1195	121	468	1257
  St. Dev.	3.6	17.43	51.39	8.35	33.23	90.5
  %CV	3.30	3.89	4.30	6.90	7.10	7.19

• ELISA Kit Component:
  • Lyophilized recombinant rat BDNF standard: 10 ng/tube x 2
  • Anti-rat BDNF Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-rat BDNF antibody: 130 uL, dilution 1:100
  • Antibody diluent buffer: 12 mL
  • Avidin-Biotin-Peroxidase Complex (ABC): 130 uL, dilution 1:100
  • ABC diluent buffer: 12 mL
  • TMB color developing agent: 10 mL
  • TMB stop solution: 10 mL
• Additional Information: Range: 31.2pg/ml-2000pg/ml
  Principle: Aviva's rat BDNF ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Rat BDNF specific-specific polyclonal antibodies were precoated onto 96-well plates. The rat specific detection monoclonal antibodies were biotinylated. The test samples and biotinylated detection antibodies were added to the wells subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the rat BDNF amount of sample captured in plate.
  Notes: 1. To inspect the validity of experiment operation and the appropriateness of sample dilution proportion, pilot
  experiment using standards and a small number of samples is recommended.
  2. The TMB Color Developing agent is colorless and transparent before using, contact us freely if it is not the case.
  3. Before using the Kit, spin tubes and bring down all components to the bottom of tubes.
  4. Duplicate well assay is recommended for both standard and sample testing.
  5. Don't let 96-well plate dry, for dry plate will inactivate active components on plate.
  6. Don't reuse tips and tubes to avoid cross contamination.
  7. To avoid to use the reagents from different batches together.
  8. In order to avoid marginal effect of plate incubation due to temperature difference ( reaction may be stronger in the
  marginal wells), it is suggested that the diluted ABC and TMB solution will be pre-warmed in 37C for 30 min before
  using.
  Background: Brain-derived neurotrophic factor (BDNF) is a prosurvival factor induced by cortical neurons that is necessary for survival of striatal neurons in the brain. It is a secreted protein with the molecular weight of 27.8kDa, consisting of 247 amino acids. It is known to promote neuronal survival and differentiation. BDNF shares substantial amino acid sequence identity with nerve growth factor (NGF). BDNF and neurotrophin-3 (NT-3) are two recently cloned neurotrophic factors that are homologous to NGF. mRNA products of the BDNF and NT-3 genes are detected in the adult human brain, suggesting that these proteins are involved in the maintenance of the adult nervous system.1 BDNF and other neurotrophins are critically involved in long-term potentiation (LTP). BDNF-mediated LTP is induced postsynaptically.2 BDNF has trophic effects on serotonergic (5-HT) neurons in the central nervous system.3 BDNF has an essential maintenance function in the regulation of anxiety-related behavior and in food intake through central mediators in both the basal and fasted state.4 It plays a role in treating breathing disorders such as respiratory insufficiency after spinal injury.5
• Reconstitution and Storage: Store at 4C for 6 months, at -20C for 12 months. Avoid multiple freeze-thaw cycles (Shipped with wet ice.)
• Sample Type: Cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA, citrate).
• Sensitivity: <15 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine.
• Reference: 1. Jones, K. R.; Reichardt, L. F. Molecular cloning of a human gene that is a member of the nerve growth factor family.
  Proc. Nat. Acad. Sci. 87: 8060-8064, 1990.
  2. Kovalchuk, Y.; Hanse, E.; Kafitz, K. W.; Konnerth, A. Postsynaptic induction of BDNF-mediated long-term
  potentiation. Science 295: 1729-1734, 2002.
  3. Lyons, W. E.; Mamounas, L. A.; Ricaurte, G. A; Coppola, V.; Reid, S. W.; Bora, S. H.; Wihler, C.; Koliatsos, V. E.;
  Tessarollo, L. Brain-derived neurotrophic factor-deficient mice develop aggressiveness and hyperphagia in
  conjunction with brain serotonergic abnormalities. Proc. Nat. Acad. Sci. 96: 15239-15244, 1999.
  4. Rios, M.; Fan, G.; Fekete, C.; Kelly, J.; Bates, B.; Kuehn, R.; Lechan, R. M.; Jaenisch, R. Conditional deletion of
  brain-derived neurotrophic factor in the postnatal brain leads to obesity and hyperactivity. Molec. Endocr. 15:
  1748-1757, 2001.
  5. Baker-Herman, T. L.; Fuller, D. D.; Bavis, R. W.; Zabka, A. G.; Golder, F. J.; Doperalski, N. J.; Johnson, R. A.;
  Watters, J. J.; Mitchell, G. S. BDNF is necessary and sufficient for spinal respiratory plasticity following intermittent
  hypoxia. Nature Neurosci. 7: 48-55, 2004.
• Specificity: Natural and recombinant rat BDNF
• Immunogen: Expression system for standard: NS0; Immunogen sequence: H129-R247
• Description: For quantitative detection of rat BDNF in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA, citrate).

Target Info

• Gene Symbol: BDNF
• Gene Full Name: brain-derived neurotrophic factor
• Alias Symbols: Abrineurin, ANON2, Brain derived neurotrophic factor, BULN2, MGC34632, Neurotrophin
• NCBI Gene Id: 24225
• Protein Name: Brain-derived neurotrophic factor
• Description of Target: Brain-derived neurotrophic factor(BDNF) is a prosurvival factor induced by cortical neurons that is necessary for survival of striatal neurons in the brain. It is a secreted protein with the molecular weight of 27.8kDa, consisting of 247 amino acids. It is known to promote neuronal survival and differentiation. BDNF shares substantial amino acid sequence identity with nerve growth factor(NGF). BDNF and neurotrophin-3(NT-3) are two recently cloned neurotrophic factors that are homologous to NGF. mRNA products of the BDNF and NT-3 genes are detected in the adult human brain, suggesting that these proteins are involved in the maintenance of the adult nervous system. BDNF and other neurotrophins are critically involved in long-term potentiation(LTP). BDNF-mediated LTP is induced postsynaptically. BDNF has trophic effects on serotonergic(5-HT) neurons in the central nervous system. BDNF has an essential maintenance function in the regulation of anxiety-related behavior and in food intake through central mediators in both the basal and fasted state. It plays a role in treating breathing disorders such as respiratory insufficiency after spinal injury.
• Uniprot ID: P23363
• Protein Accession #: NP_001257559.1
• Nucleotide Accession #: NM_001270630.1