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Product Info Summary
| SKU: | EK0995 |
|---|---|
| Size: | 96 wells/kit, with removable strips. |
| Reactive Species: | Human |
| Application: | ELISA |
| Sample Types: | cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). |
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Product info
Product Name
Human ALCAM/CD166 ELISA Kit PicoKine®
View all ALCAM/CD166 ELISA kits
SKU/Catalog Number
EK0995
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Human ALCAM/Cd166 ELISA Kit PicoKine™ (96 Tests). Quantitate Human ALCAM in cell culture supernatants, cell lysates, 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 ALCAM/CD166 ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0995)
Clonality of Antibodies
See Datasheet for details
Immunogen
Expression system for standard: NS0; Immunogen sequence: W28-A526
Sensitivity
<10 pg/ml
Assay Range
312 pg/ml - 20,000 pg/ml
Cross Reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK0995 is reactive to ALCAM in Human samples
Validated Sample Types
cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).
Application Guarantee
EK0995 is guaranteed for ELISA in Human by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of ALCAM/CD166
CD166 antigen is a 100-105 kD typeI transmembrane glycoprotein that is a member of the immunoglobulin superfamily of proteins. In humans it is encoded by the ALCAM gene. The gene was mapped to human chromosome 3q13.1-q13.2 by fluorescence in situ hybridization of cDNA probes to metaphase chromosomes. It is also called CD166 (cluster of differentiation 166), MEMD, SC-1/DM-GRASP/BEN in the chicken, and KG-CAM in the rat. It is expressed on activated T cells, activated monocytes, epithelial cells, fibroblasts, neurons, melanoma cells, and also in sweat and sebaceous glands. CD166 protein expression is reported to be upregulated in a cell line deriving from a metastasizing melanoma. CD166 plays an important role in mediating adhesion interactions between thymic epithelial cells and CD6+ cells during intrathymic T cell development. Recently, CD166 has also been used as a potential cancer stem cell marker.
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK0995-CAP | Anti-Human ALCAM Pre-coated 96-well strip microplate | 1 |
| EK0995-ST | Human ALCAM Standard | 2 vials, 20 ng/tube |
| EK0995-DA | Human ALCAM 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.
- 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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Data Examples, Quality Control Data & Sample Dilution
Validation Standard Curve O.D. At 450nm
| Concentration (pg/ml) | 0 | 312 | 625 | 1250 | 2500 | 5000 | 10000 | 20000 |
| O.D. | 0.094 | 0.276 | 0.390 | 0.635 | 1.148 | 1.589 | 1.994 | 2.274 |
Data Example Images
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Human ALCAM PicoKine ELISA Kit standard curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect ALCAM/CD166, Dilution ratio of 1:1, concentration in serum is around 2.5ng/ml.
Some articles we found to cite concentrations of ALCAM/CD166 in samples: 22475274 (Pubmed IDs).
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Human ALCAM/CD166 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 Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 16 | 16 | 16 | 24 | 24 | 24 |
| Mean (pg/ml) | 648 | 1967 | 8283 | 688 | 2156 | 8924 |
| Standard deviation | 37.58 | 145.55 | 563.24 | 48.16 | 198.35 | 651.45 |
| CV (%) | 5.8% | 7.4% | 6.8% | 7% | 9.2% | 7.3% |
Reproducibility
We ensure reproducibility by testing three samples with differing concentrations of ALCAM/CD166 in ELISA kits from four different production batches/lots.
| Lots | Lot 1 (pg/ml) | Lot 2 (pg/ml) | Lot 3 (pg/ml) | Lot 4 (pg/ml) | Mean (pg/ml) | Standard Deviation | CV (%) |
|---|---|---|---|---|---|---|---|
| Sample 1 | 648 | 630 | 693 | 657 | 657 | 22.94 | 3.4% |
| Sample 2 | 1967 | 2006 | 1987 | 2137 | 2024 | 66.54 | 3.2% |
| Sample 3 | 8283 | 8482 | 8134 | 7570 | 8117 | 339.21 | 4.1% |
Protein Target Info & Infographic
Gene/Protein Information For ALCAM (Source: Uniprot.Org, NCBI)
Uniprot ID
Q13740
Gene ID
214
Gene Name
ALCAM
Full Name
CD166 antigen
Weight
65.102kDa
Alternative Names
activated leucocyte cell adhesion molecule; activated leukocyte cell adhesion moleculeFLJ38514; ALCAM; CD166 antigen; CD166; MEMDMGC71733 ALCAM CD166, MEMD activated leukocyte cell adhesion molecule CD166 antigen
*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 ALCAM, check out the ALCAM 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 ALCAM: 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]
Specific Publications For Human ALCAM/CD166 ELISA Kit PicoKine® (EK0995)
No publications found for EK0995
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Recommended Resources
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Customer Reviews
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Customer Q&As
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7 Customer Q&As for Human ALCAM/CD166 ELISA Kit PicoKine®
Question
Q: for how much duration can samples (cell cultures, serum, and plasma) be stored and still be stable for analysis CD166 using the EK0995 Human CD166 Picokine® ELISA Kit?
Verified Customer
Verified customer
Asked: 2020-11-06
Answer
A: Boster Bio does not evaluate sample stability. Variations in sample collection, processing, and storage may change the stabilityof samples. It is recommend to assay sample immediately after collection when possible, or aliquot into single use volumes and store samples frozen. avoid repetitive freeze-thaw cycles with the stored samples in order to limit protein degradation.
Boster Scientific Support
Answered: 2020-11-06
Question
Q: how to proceed with the analysis of ELISA data? I have performed CD166 level in cell lysates.
A. Scott
Verified customer
Asked: 2020-04-14
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: 2020-04-14
Question
Q: What is the optimal O.D. value for CD166 ELISA kit? I performed your CD166 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 CD166 even though the O.D. values are not very high?
Verified Customer
Verified customer
Asked: 2019-08-20
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. a point of focus should be is whether your sample O.D. values are statistically significantly higher than your blank values. considering your assay, 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 proportion 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 CD166 ELISA kit to have sensitivity of 10pg/ml, that means the minimum amount of CD166 that can be declared/interpreted as positive by the above standard is 10pg/ml.
Boster Scientific Support
Answered: 2019-08-20
Question
Q: Are Boster Bio recombinant proteins and antibodies sterile?
B. Wu
Verified customer
Asked: 2019-08-08
Answer
A: although the vials are bottled using aseptic techniques, heat-treated vials, and sterile stock solutions, they are not considered or guaranteed to be sterile. If sterile material is needed for an experiment, the material can be filtered through a 0.2 micron filter designed for use with biological fluids.
Boster Scientific Support
Answered: 2019-08-08
Question
Q: how to thaw whole blood sample for CD166 ELISA after freezing?
Verified Customer
Verified customer
Asked: 2018-06-17
Answer
A: do not freeze and thaw whole blood. erythrocytes are fragile and, if frozen and thawed, will undergo hemolysis rendering the samples useless. To keep your blood samples to test CD166 for a later time, you should let the blood clot in glass tubes and collect the serum to freeze for later use.
Boster Scientific Support
Answered: 2018-06-17
Question
Q: we need your suggestion regarding the dilution ratio of serum samples for detection of CD166 in Human cell lysates? I am trying to measure a few analytes and it requires 100ul of diluted samples for each well. We have low serum volumes so we like to dilute as much as possible.
B. Tiwari
Verified customer
Asked: 2018-05-19
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 CD166 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: 2018-05-19
Question
Q: if the enzyme conjugated CD166 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 CD166 antigen?
Verified Customer
Verified customer
Asked: 2017-02-15
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: 2017-02-15
