Human DAND5/COCO ELISA Kit PicoKine®

COCO/DAND5 ELISA kit for Human

Human DAND5/COCO ELISA Kit PicoKine™ (96 Tests). Quantitate Human DAND5 in cell culture supernatants, serum and plasma (heparin, EDTA). Sensitivity: 10pg/ml.

Product Info Summary

SKU: EK1658
Size: 96 wells/kit, with removable strips.
Reactive Species: Human
Application: ELISA
Sample Types: cell culture supernatants, serum and plasma (heparin, EDTA).

Product Name

Human DAND5/COCO ELISA Kit PicoKine®

View all COCO/DAND5 ELISA kits

SKU/Catalog Number

EK1658

Size

96 wells/kit, with removable strips.

*Question: How many samples can I assay/run in this kit?

Description

Human DAND5/COCO ELISA Kit PicoKine™ (96 Tests). Quantitate Human DAND5 in cell culture supernatants, serum and plasma (heparin, EDTA). Sensitivity: 10pg/ml.

Storage & Handling

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Cite This Product

Human DAND5/COCO ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1658)

Clonality of Antibodies

See Datasheet for details

Immunogen

Expression system for standard: E.coli; Immunogen sequence: R23-A189

Sensitivity

<10 pg/ml

Assay Range

62.5 pg/ml - 4,000 pg/ml

Standard Dilution Instructions

serial dilution instructions image

Add 100ul of sample diluent in well #2-#8. Add 200ul standard stock solution to well #1, and serial dilute for well #2-#7 to make a standard curve row. Leave well #8 as blank See datasheet of EK1658 for more details

Cross-reactivity

There is no detectable cross-reactivity with other relevant proteins.

Reactive Species

EK1658 is reactive to Dand5 in Human samples

Validated Sample Types

cell culture supernatants, serum and plasma (heparin, EDTA).

Application Guarantee

EK1658 is guaranteed for ELISA in Human by Boster Guarantee

See how Boster Bio validate our ELISA kits: ELISA Validation Information

Background of COCO/DAND5

This gene encodes a member of the BMP (bone morphogenic protein) antagonist family. Like BMPs, BMP antagonists contain cystine knots and typically form homo- and heterodimers. The CAN (cerberus and dan) subfamily of BMP antagonists, to which this gene belongs, is characterized by a C-terminal cystine knot with an eight-membered ring. The antagonistic effect of the secreted protein encoded by this gene is likely due to its direct binding to BMP proteins. As an antagonist of BMP, this gene may play a role in regulating organogenesis, body patterning, and tissue differentiation. In mouse, this protein has been shown to bind Nodal and to inhibit the Nodal signaling pathway which patterns left/right body asymmetry.

Kit Components

Catalog Number Description Quantity
EK1658-CAP Anti-Human Dand5 Pre-coated 96-well strip microplate 1
EK1658-ST Human Dand5 Standard 2 vials, 10 ng/tube
EK1658-DA Human Dand5 Biotinylated antibody (100x) 100ul
AR1103 Avidin-Biotin-Peroxidase Complex (100x) 100ul
AR1106-1 Sample Diluent 30ml
AR1106-2 Antibody Diluent 12ml
AR1106-3 Avidin-Biotin-Peroxidase Diluent 12ml
AR1104 Color Developing Reagent (TMB) 10ml
AR1105 Stop Solution 10ml
AR1106-5 Wash Buffer (25x) 20ml
PLA-SEA Adhesive plate sealers 4

*The kit components are not available for individual purchase.

Materials Required But Not Included In Kit

  • Microplate Reader capable of reading absorbance at 450nm.
  • Incubator.
  • Automated plate washer (optional).
  • Pipettes and pipette tips capable of precisely dispensing 0.5 µl through 1 ml volumes of aqueous solutions.
  • Multichannel pipettes are recommended for large amount of samples.
  • Deionized or distilled water.
  • 500ml graduated cylinders.
  • Test tubes for dilution.

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Validation Standard Curve O.D. At 450nm

Concentration (pg/ml)062.5125250500100020004000
O.D.0.0630.1170.1770.2740.4970.841.4972.174

Data Example Images

Recommended Sample Dilution Ratios

According to our internal validation assays using this ELISA kit, to detect COCO/DAND5, Dilution ratio of 1:1, concentration in serum and plasma is less than the lowest standard, 62.5 pg/ml..

Intra Assay Consistency & Inter Assay Consistency

We measured random samples of Human DAND5/COCO ELISA Kit PicoKine® within the same batch/lot to ensure the consistency of the kits' performances. ELISA intra assay consistency is measured using wells from the same plate/assay kit. ELISA inter assay consistency is measured using wells from different plates from the same batch production/lot.

Intra-Assay PrecisionInter-Assay Precision
Sample123123
n161616242424
Mean (pg/ml)16463618891516431700
Standard deviation10.6643.8810210.5750.7996.9
CV (%)6.5%6.9%5.4%7%7.9%5.7%

Reproducibility

We ensure reproducibility by testing three samples with differing concentrations of COCO/DAND5 in ELISA kits from four different production batches/lots.

LotsLot 1 (pg/ml)Lot 2 (pg/ml)Lot 3 (pg/ml)Lot 4 (pg/ml)Mean (pg/ml)Standard DeviationCV (%)
Sample 11641651551721646.043.6%
Sample 263666966956863541.236.4%
Sample 31889182718781962188948.22.5%
*number of samples for each test n=16.

Gene/Protein Information For Dand5 (Source: Uniprot.Org, NCBI)

Gene Name

Dand5

Full Name

DAN domain family member 5

Weight

19.772kDa

Superfamily

DAN family

Alternative Names

CER2CRL2; Cerberus 2; Cerberus like-2; cerberus-like 2; Cerberus-like protein 2; CERL2; cerl-2; CKTSF1B3; CKTSF1B3MGC126849; COCO; Cysteine knot superfamily 1, BMP antagonist 3; DAN domain family member 5; DAN domain family, member 5; DAND5; Dante; DTE; FLJ38607; GREM3; GREM3cerberus 2; gremlin-3; SP1 Dand5|Cer, Cerl, Cerl-2, Cerl2, Cerr2, Dte, GREM, GREM3, co, coco|DAN domain family member 5, BMP antagonist|DAN domain family member 5|DAN domain family, member 5|cerberus-like protein 2|dante

*if product is indicated to react with multiple species, protein info is based on the gene entry specified above in "species".

For more info on Dand5, check out the Dand5 Infographic

Dand5 infographic

We have 30,000+ of these available, one for each gene! check them out.

In this infographic you will see the following information for Dand5: database IDs, super-family, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact us [email protected].

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9 Customer Q&As for Human DAND5/COCO ELISA Kit PicoKine®

Question

Q: for how much duration can samples (cell cultures, serum, and plasma) be stored and still be stable for measurement COCO using the EK1658 Human COCO Picokine® ELISA Kit?

Verified Customer

Verified customer

Asked: 2020-10-20

Answer

A: Boster Bio does not evaluate sample stability. Variations in sample collection, processing, and storage may affect the stabilityof samples. It is recommend to assay sample on-stat collection when possible, or aliquot into single use volumes and store samples frozen. avoid repeat freeze-thaw cycles with the stored samples to limit protein degradation.

Boster Scientific Support

Answered: 2020-10-20

Question

Q: how to analyze ELISA data? I have obtained COCO level in serum.

Verified Customer

Verified customer

Asked: 2020-08-24

Answer

A: we recommend you this article on ELISA data analysis. bosterbio.com/ELISA-data-analysis-instructions. Boster also provides a free tool that you can use to analyze ELISA data. bosterbio.com/biology-research-tools/ELISA-data-analysis-online

Boster Scientific Support

Answered: 2020-08-24

Question

Q: is there any online tool I can use to streamline the data analysis for my ELISA results?

M. Allen

Verified customer

Asked: 2019-11-21

Answer

A: We have a web based ELISA curve fitting (4pl) and data analysis tool. Please give it a try: bosterbio.com/biology-research-tools/ELISA-data-analysis-online. You can also consult our article on ELISA data analysis: bosterbio.com/ELISA-data-analysis-instructions

Boster Scientific Support

Answered: 2019-11-21

Question

Q: if the enzyme conjugated COCO antibodies are mixed with the substrate, will that change the substrate into the enzymatic reaction product? Or the enzyme function is only activated when the antibody is attached with the COCO antigen?

C. Sunder

Verified customer

Asked: 2019-10-01

Answer

A: The enzyme is always active. Avoid contaminating the substrate with enzyme prior to the incubation otherwise it compromises the assay with false positive signal.

Boster Scientific Support

Answered: 2019-10-01

Question

Q: how many samples can be assayed in a Picokine® ELISA Kit?

Verified Customer

Verified customer

Asked: 2019-09-01

Answer

A: The Picokine® ELISA Kits will generally run a 7-point standard curve, non-specific binding wells, and 39 samples in duplicate. this may vary slightly by kit so please refer to each datasheet for details.

Boster Scientific Support

Answered: 2019-09-01

Question

Q: we need your suggestion regarding the dilution ratio of serum samples for detection of COCO in Human serum? I am trying to measure a few analytes and it requires 100ul of diluted samples for each well. We have low serum quantitys so we like to dilute as much as possible.

Verified Customer

Verified customer

Asked: 2019-07-29

Answer

A: without knowing the physiological or pathological context of your samples we cannot recommend a dilution ratio without performing a pilot test with your samples. Here is how you can perform a pilot study on your own: perform a serial dilution of your samples on the COCO ELISA kit to make sure you have a linear ascending curve followed by a plateau, which signifies the samples saturating the detection limit of the kit. Then you can pick the dilution ratios from samples in the linear part of the curve as your experimental dilution ratio.
If you are interested in using our ELISA service, you can also send us your sample and we will take care of everything for you. You can check our service details here: bosterbio.com/services/assay-services/ELISA-testing-service
Since you mentioned you have limited samples, our cost effective multiplex ELISA service would fit perfectly for your needs, where we can generate dozens of data points using as little as 25ul sample volume. Information on this service is also in the above link.

Boster Scientific Support

Answered: 2019-07-29

Question

Q: What is the optimal O.D. value for COCO ELISA kit? I performed your COCO ELISA on serum samples. For my positive control, I received an O.D. value of 0.826, while my negative control received a value of 0.136. I obtained both of these controls from the ELISA kit, where your kit's typical data shows O.D. values much higher than my positive control and your background is lower. My samples O.D. values are around 0.225 and the highest is only 0.357. is it safe to say these samples contain COCO even though the O.D. values are not very high?

G. Banerjee

Verified customer

Asked: 2019-07-04

Answer

A: The absolute O.D. values may change according to incubation time. The more you incubate the higher the O.D. values are going to be. what you should focus on is whether your sample O.D. values are statistically significantly higher than your blank values. in your example, you could extend your development time in the substrate incubation step to obtain higher O.D. values, as long as your negative controls' O.D. values are not increasing faster in relation to your positive controls. typically, a sample with O.D. value 2 standard deviations higher than your negative controls can be considered positive. We calculate the sensitivity of this ELISA kit by converting cutoff O.D. value, calculated as the average of 20 negative controls plus 2 standard deviations of the 20 negative controls, into a concentration. in other words, when we claim this COCO ELISA kit to have sensitivity of 10pg/ml, that means the minimum amount of COCO that can be declared/interpreted as positive by the above standard is 10pg/ml.

Boster Scientific Support

Answered: 2019-07-04

Question

Q: is it okay to use citrate plasma as samples in Human COCO Picokine® ELISA Kit (Catalog # EK1658)?

W. Dhar

Verified customer

Asked: 2019-01-23

Answer

A: Chelating agents such as EDTA, Heparin and Citrate can attach metal ions from the functional domain of COCO causing degradation of its protein structure. COCO may be denatured as a result and may compromise the assay's measurements. The chilating sites could also be too close to the epitopes required for detection and block the antigen antibody reaction. We have tested the COCO ELISA, treating samples with a number of anticoagulants and decided that heparin or EDTA can be used for treatment of blood/plasma samples. Do not use other anticoagulents when collecting samples.

Boster Scientific Support

Answered: 2019-01-23

Question

Q: how are cell lysates prepared for use in Picokine® ELISA kits?

Verified Customer

Verified customer

Asked: 2018-03-02

Answer

A: for those Picokine® ELISAs where cell or tissue lysate is a validated sample type, sample preparation instructions for lysate are included in the product insert. Components in lysate and lysis buffer can impact immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.

Boster Scientific Support

Answered: 2018-03-02

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