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SKU:EK0307
Application:ELISA
Sensitivity:<15pg/ml
Sample Type:cell culture supernates, cell lysates, serum and plasma (heparin, EDTA, citrate).
Price: $270.00
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Overview

Product Name Human BDNF PicoKine™ ELISA Kit
Description For quantitative detection of human BDNF in cell culture supernates, cell lysates, serum and plasma (heparin, EDTA, citrate).
Cite This Product Human BDNF PicoKine™ ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0307)
Replacement Item This antibody may replace the following items: sc-546|sc-33905|sc-33904|sc-20981 from Santa Cruz Biotechnology.
Validated Species Human
Predicted Species Bovine, Horse, Monkey, Rabbit

*This antibody is predicted to react with the above species based on antigen sequence similarities. Our Boster Guarantee covers the use of this product with the above species.

Application ELISA

*Our Boster Guarantee covers the use of this product in the above tested applications.

**For positive and negative control design, consult "Tissue specificity" under Protein Target Info.

Immunogen Expression system for standard: sf21; Immunogen sequence: H129-R247
Cross Reactivity There is no detectable cross-reactivity with other relevant proteins.
Pack Size 96wells/kit

Properties

Sensitivity <15pg/ml
*Sensitivity, or Lower Limit of Detection (LLD), is the minimum level of target protein the ELISA assay can detect. We measure 20 blank wells and if the O.D. value is 2 standard deviations higher than the blanks' average O.D. the sample can be deemed positive.
Assay Range 31.2pg/ml-2000pg/ml
*This assay range is determined using common samples. For samples with low target protein concentrations, users can adjust the standard curve to extend the lower limit of assay range.
Sample Type cell culture supernates, cell lysates, serum and plasma (heparin, EDTA, citrate).

*The above listed samples are the ones valided with the assay. If you do not see your sample of interest listed, as long as there is enough level of target protein present in the sample, this Picokine™ ELISA kit should detect it.
**For protocol and tips regarding preparing your sample of interest, please check our ELISA sample preparation guide.
Capture Antibody monoclonal antibody from mouse
Detection Antibody polyclonal antibody from goat
Natural Levels Some research articles suggesting the natural levels of BDNF are listed below:
Storage Store at 4˚C for 6 months, at -20˚C for 12 months. Avoid multiple freeze-thaw cycles(Shipped with wet ice.)

Kit Components

Catalog numberDescriptionQuantity
EK0307-CAP96-well plate precoated with anti-Human BDNF antibody1
EK0307-STlyophilized recombinant Human BDNF standard10ng/tubex2
EK0307-DAbiotinylated anti-Human BDNF antibody130ul(dilution 1:100)
AR1103Avidin-Biotin-Peroxidase Complex(ABC)130ul(dilution 1:100)
AR1106-1sample diluent buffer30ml
AR1106-2antibody diluent buffer12ml
AR1106-3ABC diluent buffer12ml
AR1104TMB color developing agent10ml
AR1105TMB stop solution10ml
PLA-SEAAdhesive cover4
AR0030-EPBS washing bufferPowder for 1000ml

Washing buffer Preparation: Disolve AR0030-E to 1000ml distilled water and adjust pH to 7.2~7.6. Finally, adjust the total volume to 1L.

*Additional components can be purchased. If you need extra of the above components please order them together to avoid additional shipping charges.

See Prices For Extra Components

Material Required But Not Provided

  • 1. Microplate reader in standard size.
  • 2. Automated plate washer.
  • 3. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection.
  • 4. Clean tubes and Eppendorf tubes.
*Reagents needed for sample collection and preparation are not included in the kit.

Protein Target Info (Source: Uniprot.org)

You can check the tissue specificity below for information on selecting positive and negative control.

Gene Name BDNF
Protein Name Brain-derived neurotrophic factor
Molecular Weight 27818 MW
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. The versatility of BDNF is emphasized by its contribution to a range of adaptive neuronal responses including long-term potentiation (LTP), long-term depression (LTD), certain forms of short-term synaptic plasticity, as well as homeostatic regulation of intrinsic neuronal excitability. .
Tissue Specificity Brain. Highly expressed in hippocampus, amygdala, cerebral cortex and cerebellum. Also expressed in heart, lung, skeletal muscle, testis, prostate and placenta. .
Sequence Similarities Belongs to the NGF-beta family.
Subcellular Localization Secreted.
Uniprot ID P23560
Alternative Names Brain-derived neurotrophic factor;BDNF;Abrineurin;BDNF;
Research Areas |neuroscience|neurology process|growth and development|neurotrophins| neuroscience|neurodegenerative disease|huntington's disease|alzheimer's disease| cardiovascular|atherosclerosis|diabetes associated|thrombosis| metabolism|types of disease|diabetes|obesity|
*if product is indicated to react with multiple species, protein info is based on the human gene.

Background for Brain-derived neurotrophic factor

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. The mature form of BDNF is identical in all mammals examined, and the gene encoding human BDNF to chromosome 11, band p13.

Human BDNF PicoKine™ ELISA Kit Images

Click the images to enlarge.

Human BDNF PicoKine™ ELISA Kit
Human BDNF PicoKine ELISA Kit standard curve

Intra/Inter Assay Precision

>
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n161616242424
Mean(pg/ml)11342512781975941306
Standard deviation3.9617.8562.6214.1844.55103.17
CV(%)3.54.24.97.27.57.9

Typical Data Obtained from Human BDNF PicoKine™ ELISA Kit

(TMB reaction incubate at 37°C for 25-30min)

Concentration(pg/ml)031.262.512525050010002000
O.D.0.0630.0950.1330.2160.3580.7201.4052.298

*The typical data is obtained from Boster's internal QC result and for reference only. It may differ from the lab test results of the end users. It is more important that the user's lab test results reflect the same linearity demonstrated in the typical data than achieving exactly the same O.D. values.

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Publications

Schulze, J., Kaiser, O., Paasche, G., Lamm, H., Pich, A., Hoffmann, A.,..., & Warnecke, A. (2017). Effect of hyperbaric oxygen on BDNF-release and neuroprotection: Investigations with human mesenchymal stem cells and genetically modified NIH3T3 fibroblasts as putative cell therapeutics. PLoS One, 12(5), e0178182. doi: 10.1371/journal.pone.0178182
Burnout and cognitive impairment: Associated with serum BDNF in a Chinese Han population S.C. Hea, , , Y.Y. Zhangb, J.Y. Zhana, C. Wanga, X.D. Dub, G.Z. Yinb, B. Caoc, Y.P. Ningd, J.C. Soaresc, X.Y. Zhangc, e, Burnout and cognitive impairment: Associated with serum BDNF in a Chinese Han population
Ankur Jindala, Radhakrishnan Mahesha, Shvetank Bhatta, Dilip Pandeyb Neurochemistry International Available online 14 December 2016 Molecular modifications by regulating cAMP signaling and oxidant-antioxidant defence mechanisms, produce antidepressant-like effect: A possible mechanism of etazolate aftermaths of impact accelerated traumatic brain injury in rat model
Shi Y, You J, Yuan Y, Zhang X, Li H, Hou G. Psychiatry Clin Neurosci. 2010 Jun;64(3):249-54. Doi: 10.1111/J.1440-1819.2010.02074.X. Plasma Bdnf And Tpa Are Associated With Late-Onset Geriatric Depression.
Teche Sp, Nuernberg Gl, Sordi Ao, De Souza Lh, Remy L, Ceres??r Km, Rocha Ns. Psychiatr Q. 2013 Dec;84(4):485-97. Doi: 10.1007/S11126-013-9261-7. Measurement Methods Of Bdnf Levels In Major Depression: A Qualitative Systematic Review Of Clinical Tr...
Zakharyan R, Boyajyan A. Clin Biochem. 2014 Aug;47(12):1052-5. Doi: 10.1016/J.Clinbiochem.2014.03.021. Epub 2014 Apr 5. Brain-Derived Neurotrophic Factor Blood Levels Are Decreased In Schizophrenia Patients And Associate With Rs6265 Genotypes.

FAQs

Q: The protocol says to use neutral PBS and provides a recipe. Could I use neutral PBS made with KCl or KH2PO4 (common constituents for most PBS recipes) instead? Keywords: protocol, alternative buffer, applications, reagent recipe
A: Yes, it is ok to use common PBS recipes. We have tried many types of buffers with common constituents, and no significant difference was observed whatsoever.
Q: What is the expression system for the standards? Do the standards contain carrier protein? Keywords: product immunogen information, antigen sequence, expression standard
A: The expression system of the standards can be found in the product page under ""immunogen"". It tells you both the expression system and the sequence information. The standards do contain carrier protein. The carrier protein is BSA unless the target protein is also present in BSA--in such cases please contact us for details of the carrier protein.
Q: Why are my O.D. values different than your values on the datasheet? Keyword: troubleshooting, reference
A: We detected the kit in our lab and got our values on the datasheet before delivery, but protein activity will decrease as time goes on, so you may get lower O.D. values than ours. However, it should still be in a reasonable range and the standards can be used to calculate sample values. Good linearity of curve is more important than the actual numerical value.
Q: On the ELISA kit datasheet it says "HRP substrate TMB was used to visualize HRP enzymatic reaction." What is HRP and which part of the kit contains HRP? Keyword: kit components, reagents, protocol
A: It means Avidin-Biotin-Peroxidase Complex(AR1103) and you could find it in Kit Components.
Q: Do you offer any ELISA kits that can work with whole blood samples instead of plasma or urine? Keyword: applications
A: We test serum and plasma routinely, and there is very little difference between serum, plasma and whole blood. The whole blood also contains proteins we need to test. The kit can be used to test whole blood in theory.
Q: Does silicic acid (formula SiOH4) affect the results of the ELISA assay? Keyword: protocol, reagents
A: The acid may affect the binding of antigen to antibody, it is not recommended to use. That being said, it is unlikely to affect the reaction if the solution remains an overall neutral environment.
Q: Is there lot-to-lot variation of the ELISA kits? What are Boster's general services when I have questions about your kits? Keyword: technical support, help, customer service
A: The NIBSC/WHO 1st International Standard is evaluated, we always test our kits before delivery, and customers can find the test result on the protocol. If the customer needs technical support from us to analyze their data, please contact us and we ask that you provide us the following information: the lot# and production date, and when did the customer detect the kit.
Q: The results of my standards do not look correct, what could be the problem? Keyword: troubleshoot, protocol
A: Double check the protocol for plate washing. It should be examined on three aspects: 1) Did you make the washing buffer correctly? 2) How long did you let the buffer stay in wells? 3) What was the volume of the washing buffer added to each well? In addition, pay attention when adding sample to avoid contamination. And we suggest testing the standards again. Values of standards at low concentration are more affected than that at higher concentration, so customer can still get expected values at high concentration even if there is an error. If problems persist, please contact technical support with the catalog and lot number of your product.
Q: What is the well depth of the 96 well plates for the ELISA kits? Keyword: well capacity, product size
A: The well depth is 300ul, and the max capacity is 350ul. The height of the well is 1.1 cm, and the internal size is 1 cm.
Q: The kit does not include wash solution, what should I do? Keyword: wash buffer
A: Our Elisa kits do not come with wash solution, but we have included information about how to make washing buffer on the datasheet. Please refer to the "Material Required But Not Provided" section. We also offer washing buffer for sale separately (Phosphate Buffered Saline Powder SKU: AR0030).
Q: For your ELISA kits, are the capture and detection antibodies polyclonals or monoclonals? Keyword: antibody clonality
A: This information can be found for each kit under the "Properties" section, and you can find the immunogen sequence information in the "Overview" section.
Q: Do your ELISA kits come with sealants or plate covers? Keyword: storage, sequential use
A: The kit can be used within a month sequentially if it's opened and stored at 4 degree. There is no need to use sealants, the plate can be packaged with aluminum foil bag, and for other reagents keep bottle tightly closed.
Q: What is the concentration of Sodium Azide in your Elisa kits? Keyword: preservative
A: Our Elisa kits contain 0.02% Sodium Azide. All of the components except TMB colour developing reagent and stop solution contains 0.02% Sodium azide as the preservative.
Q: Are there positive and negative controls available for my ELISA kit? Keyword: positive control, negative control
A: We can provide a recombinant protein as a control. It costs $50 per control and takes 2 weeks to manufacture. We cannot provide a positive or negative control in serum.
Q: Why do I get positive results for my knockout (KO) model when used as a control?
A: The knockout (KO) model may contain truncated forms of the target protein which can be detected by the ELISA assay.
Q: How long do I soak my plate in the wash buffer?
A: The plate should soak in 0.3mL PBS or TBS wash buffer for at least 1-2 minutes in an automated wash station.
Q: Can I use Tween in my wash buffer?
A: While it is not recommended to use Tween in your wash buffer, small amounts (<0.1% concentration) may decrease background due to insufficient blocking.
Q: What plate type do I use to set up the microplate reader?
A: Our plates are made with the Corning costar plates similar to the DNA-BIND 96 -well plates.
Q: Will this ELISA kit work on saliva or nasal washes?
A: We have not validated this kit on saliva or nasal washes.It can be detected if the concentration of BDNF in saliva or nasal wash samples are in the detection range of the kit.
Q: My samples have highly concentrated levels of BDNF and require more than normal dilution. Can you provide any extra (~ 90 mL) sample diluent? Do you sell the diluent separately?
A: Yes, we sell the diluent separately (Boster Catalog# AR1106-1). In fact, the sample diluent buffer in the kit is generally enough. For instance, if the dilution ratio is 1:20000, it is not necessary to dilute the samples at 1:20000 in just one step. Instead, we suggest you sequentially dilute the samples: 1:100, 1.200, etc. That way, you can use less diluent buffer.
Q: I am interested in performing Western Blot (WB) using the antibodies from your human and mouse BDNF ELISA kits. Is it possible to order the primary antibody (the capture antibody) that is pre-coated in the wells for the sandwich ELISA? (In this case that would be the monoclonal antibody from rat for the mouse kit, and the monoclonal antibody from mouse for the human kit). Is it also possible to order your secondary antibody which is the biotinylated detection polyclonal antibody from goat specific for BDNF?
A: The antibodies used in those ELISA kits are not fully validated on WB. Also, since these antibodies are made from recombinant proteins, they are often less reactive to retrieved antigens in WB or IHC due to the lack of tertiary structure in the retrieved antigens. If you want to test human and mouse BDNF with WB, please use our anti-BDNF antibody (PB9075). Further, you can separately buy our secondary antibodies.
Q: Which isoform (mature or precursor) does this ELISA assay detect? I am also wondering if the dimer form of mature BDNF can be detection by ELISA.
A: The assay recognizes both of mature and precursor isoforms of BDNF and can detect the dimer form.
Q: What should I use for negative control?
A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signal
A: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
Q: What is the normal level of this protein in my sample of interest?
A: We have reviewed literature and have gathered this information for most of our ELISA kits. You can find this information on the product page or contact us if you cannot find it. However this information is only suggestive and cannot replace pilot studies in determining the optimal sample dilution ratio.
Q: Is the plate separable? Can I use only part of the kit and save the rest for later? How many samples can I run with one kit?
A: Yes the plate is separable. There are 12 strips of 8 wells each, all removable from the plate. The amount of samples you can run depend on a few factors. In the most common ELISA set up, you will use two strips for standards, and 10 strips for samples using duplicates, which let you run up to 40 samples per kit. Contact us if you have questions regarding other situations.
Q: Does the kit contain sample preparation reagents? How do I prepare my samples?
A: Since different sample types require different reagents, we do not include them in the ELISA kit. However we do have each reagent mentioned in the file below available at very reasonable prices. Be sure to check them out. For sample preparation protocols please download the file below: https://www.bosterbio.com/media/pdf/Boster_Sample_Preparation_Protocols.pdf
Q: Can this ELISA kit work on brain tissue homogenate, cell culture supernatant, saliva, urine, serum, whole blood or any other sample type?
A: In theory the ELISA kit will work for all sample types if the target protein is present at a level that falls within the linear range of the ELISA kit detection range. We guarantee the kit to work on the sample types that we have tested. If you want dilution ratio suggestions on these sample types please contact our technical support. For sample types that we have not tested for, we suggest you run pilot experiments to decide the optimal sample dilution.
Q: Can this ELISA kit react with the pro-form of the target protein? Can this ELISA kit react with an isoform of the protein?
A: In general, unless otherwise specified, the ELISA kit is pan-specific, meaning that it will react with all different forms of the target protein if they share the majority of the target protein's sequence. The capture and detection antibodies are reactive to the entire sequence of the standard protein. You can find the sequence information of the standard protein in the "immunogen" section. For more information about the specificity of the kit for your particular experiment, please contact our techincal support.
Q: Can this ELISA kit react with human, mouse, rat or other species?
A: If the kit is reactive to another commonly used species (human mouse and or rat) we would have listed it as a separate product. If you want to check cross reactivity to a species that is not included in the 3 species listed above, please contact our technical support for more information. As a rule of thumb, if the sequences are 90%+ indentical, there is a high chance of cross reactivity for your species of interest.