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Product Info Summary
| SKU: | EK0838 |
|---|---|
| Size: | 96 wells/kit, with removable strips. |
| Reactive Species: | Human |
| Application: | ELISA |
| Sample Types: | cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). |
Product info
Product Name
Human Diablo/SMAC ELISA Kit PicoKine®
View all SMAC/Diablo ELISA kits
SKU/Catalog Number
EK0838
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Human Diablo/SMAC ELISA Kit PicoKine® (96 Tests). Quantitate Human DIABLO in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Sensitivity: 10pg/ml. The brand Picokine indicates this is a premium quality ELISA kit. Each Picokine kit delivers precise quantification, high sensitivity, and excellent reproducibility. Only our most reliable and effective kits qualify as Picokine, guaranteeing top-tier results for your assays.
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 Diablo/SMAC ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0838)
Clonality of Antibodies
See Datasheet for details
Standard Protein
Expression system for standard: E.coli; Immunogen sequence: A56-D239
Sensitivity
<10 pg/ml
Assay Range
31.2 pg/ml - 2,000 pg/ml
Standard Dilution Instructions
See datasheet of EK0838 for more details
Cross-reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK0838 is reactive to DIABLO in Human samples
Validated Sample Types
cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).
Application Guarantee
EK0838 is guaranteed for ELISA in Human by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of SMAC/Diablo
Diablo homolog, mitochondrial, also known as second mitochondria-derived activator of caspases (SMAC), is a protein that in humans is encoded by the DIABLO gene (direct IAP binding protein with low pI). It is mapped to 12q24.31. DIABLO can bind mammalian IAP homolog A (MIHA, or API3) and can also interact with MIHB, MIHC, and OpIAP, the baculoviral IAP. This gene encodes an inhibitor of apoptosis protein (IAP)-binding protein. The encoded mitochondrial protein enters the cytosol when cells undergo apoptosis, and allows activation of caspases by binding to inhibitor of apoptosis proteins. Over expression of the encoded protein sensitizes tumor cells to apoptosis. A mutation in this gene is associated with young-adult onset of nonsyndromic deafness-64.
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK0838-CAP | Anti-Human DIABLO Pre-coated 96-well strip microplate | 1 |
| EK0838-ST | Human DIABLO Standard | 2 vials, 10 ng/tube |
| EK0838-DA | Human DIABLO 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.
Data Examples, Quality Control Data & Sample Dilution
Validation Standard Curve O.D. At 450nm
| Concentration (pg/ml) | 0 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 | 2000 |
| O.D. | 0.024 | 0.049 | 0.096 | 0.193 | 0.358 | 0.773 | 1.338 | 2.094 |
Data Example Images
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Human Diablo/SMAC PicoKine ELISA Kit standard curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect SMAC/Diablo, Dilution ratio of 1:1, concentration in serum and plasma is around 60 pg/ml..
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Human Diablo/SMAC 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) | 54 | 221 | 865 | 53 | 229 | 932 |
| Standard deviation | 4.05 | 17.01 | 38.83 | 4.61 | 20.15 | 48.46 |
| CV (%) | 7.5% | 7.7% | 4.2% | 8.7% | 8.8% | 5.2% |
Reproducibility
We ensure reproducibility by testing three samples with differing concentrations of SMAC/Diablo 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 | 54 | 54 | 55 | 55 | 54 | 0.5 | 0.9% |
| Sample 2 | 221 | 218 | 222 | 235 | 224 | 6.51 | 2.9% |
| Sample 3 | 865 | 888 | 885 | 882 | 880 | 8.91 | 1% |
Protein Target Info & Infographic
Gene/Protein Information For DIABLO (Source: Uniprot.Org, NCBI)
Gene Name
DIABLO
Full Name
Diablo homolog, mitochondrial
Weight
27131 MW
Alternative Names
Diablo homolog, mitochondrial;Direct IAP-binding protein with low pI;Second mitochondria-derived activator of caspase;Smac;DIABLO;SMAC; DIABLO DFNA64, SMAC diablo IAP-binding mitochondrial protein diablo homolog, mitochondrial|diablo-like protein|direct IAP-binding protein with low pI|second mitochondria-derived activator of caspase
*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 DIABLO, check out the DIABLO 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 DIABLO: 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 Diablo/SMAC ELISA Kit PicoKine® (EK0838)
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No publications found for EK0838
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Customer Reviews
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Customer Q&As
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11 Customer Q&As for Human Diablo/SMAC ELISA Kit PicoKine®
Question
Q: Can SMAC ELISA Kits be used with tissue homogenates (or other non-validated sample types)?
B. Krishna
Verified customer
Asked: 2020-12-17
Answer
A: Unfortunately, Boster Bio has not routinely validated tissue homogenates as a sample type for ELISA kits. This does not mean that ELISA kits are not valid for other sample types than we have tested: it means further investigation is needed. One will need to perform a spike and recovery study to determine if an unvalidated sample type will work with a particular kit. To perform a spike and recovery experiment, one should divide a sample into two aliquots. In one of the aliquots, the user should spike in a known amount of the kit standard. a dilution series is performed comparing the spiked versus the unspiked sample. Generally, samples with expected recovery and linearity between 80-120% are considered acceptable. This method can be used to validate any sample type that has not been evaluated by Boster Bio. for a more detailed spike and recovery protocol, please contact technical support.
Note: acceptable ranges should be determined individually by each laboratory. Additionally, technical support can help determine if a buffer component is not compatible with a given ELISA kit. please view the Citations tab on the product webpage for peer-reviewed papers utilizing a wide range of sample types. We also have an innovator's reward program where if the user validates our ELISA kits in applications or samples previously not validated by Boster Bio or other users, and share such information with us by submit a review, we will reward the user's efforts with a free antibody or ELISA kit from our catalog. Biocompare.com will also give $20 Amazon giftcard as an additional reward, if the review is submitted there as well.
Boster Scientific Support
Answered: 2020-12-17
Question
Q: Are Boster Bio recombinant proteins and antibodies sterile?
Verified Customer
Verified customer
Asked: 2020-06-27
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 required for an experiment, the material can be filtered through a 0.2 micron filter designed for use with biological fluids.
Boster Scientific Support
Answered: 2020-06-27
Question
Q: can you recommend the dilution ratio of serum samples for detection of SMAC in Human plasma? I am trying to measure a a number of analytes and it requires 100ul of diluted samples for each well. We have limited sample quantitys so we like to dilute as much as possible.
Verified Customer
Verified customer
Asked: 2019-10-31
Answer
A: having little idea about 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 SMAC 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: 2019-10-31
Question
Q: how to proceed with the analysis of ELISA data? I measured SMAC level in plasma.
Verified Customer
Verified customer
Asked: 2019-08-13
Answer
A: please read this article on ELISA data analysis. bosterbio.com/ELISA-data-analysis-instructions. we also provide a convenient online tool that you can use to analyze ELISA data. bosterbio.com/biology-research-tools/ELISA-data-analysis-online
Boster Scientific Support
Answered: 2019-08-13
Question
Q: how much samples can be assayed in a Picokine® ELISA Kit?
C. Nelson
Verified customer
Asked: 2019-07-27
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: 2019-07-27
Question
Q: for how much duration can samples (cell cultures, serum, and plasma) be stored and still be stable for measurement SMAC using the EK0838 Human SMAC Picokine® ELISA Kit?
Verified Customer
Verified customer
Asked: 2019-03-11
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 on-stat collection when possible, or aliquot into single use volumes and store samples frozen. avoid repeat freeze-thaw cycles with the stored samples in order to limit protein degradation.
Boster Scientific Support
Answered: 2019-03-11
Question
Q: how are cell lysates prepared for use in Picokine® ELISA kits?
Verified Customer
Verified customer
Asked: 2019-02-21
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 have an affect on immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.
Boster Scientific Support
Answered: 2019-02-21
Question
Q: What is the optimal O.D. value for SMAC ELISA kit? I used your SMAC 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 SMAC even though the O.D. values are not very high?
K. Williams
Verified customer
Asked: 2019-01-24
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. in the example mentioned, 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. normally, 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 changeing 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 SMAC ELISA kit to have sensitivity of 10pg/ml, that means the minimum amount of SMAC that can be declared/interpreted as positive by the above standard is 10pg/ml.
Boster Scientific Support
Answered: 2019-01-24
Question
Q: can I use citrate plasma as samples in Human SMAC Picokine® ELISA Kit (Catalog # EK0838)?
Verified Customer
Verified customer
Asked: 2019-01-09
Answer
A: Chelating agents such as EDTA, Heparin and Citrate can sequester metal ions from the functional domain of SMAC causing degradation of its protein structure. SMAC may be denatured as a result and may compromise the assay's measurements. The chilating sites could also be too close to the epitopes a must for detection and block the antigen antibody reaction. We have tested the SMAC 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: 2019-01-09
Question
Q: if the enzyme conjugated SMAC 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 SMAC antigen?
N. Li
Verified customer
Asked: 2018-11-26
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: 2018-11-26
Question
Q: how can I thaw whole blood sample for SMAC ELISA after freezing?
H. Kumar
Verified customer
Asked: 2016-07-03
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 SMAC for a later time, you should let the blood clot in glass tubes and separate the serum to freeze for later analysis.
Boster Scientific Support
Answered: 2016-07-03
