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Product Info Summary
| SKU: | EK0827 |
|---|---|
| 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 Rage ELISA Kit PicoKine®
SKU/Catalog Number
EK0827
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Pathways/roles: RAGE (AGER) ligation by AGEs and other ligands → NF-κB activation; associated with chronic inflammatory and vascular pathologies (from product page background). Assay context: sandwich ELISA; validated sample types include cell culture supernatants, cell lysates, serum, plasma (heparin/EDTA); sensitivity <10 pg/mL; range 78–5,000 pg/mL; no detectable cross-reactivity reported. Often assessed with vascular/inflammation markers like Endothelin-1 and neutrophil enzyme MPO to profile cardiometabolic risk.
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 Rage ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0827)
Clonality of Antibodies
See Datasheet for details
Standard Protein
Expression system for standard: NS0; Immunogen sequence: Q24-A344
Capture/Detection Antibodies
The capture antibody is monoclonal antibody from mouse and the detection antibody is polyclonal antibody from goat.
Sensitivity
<10 pg/ml
Assay Range
78 pg/ml - 5,000 pg/ml
Standard Dilution Instructions
See datasheet of EK0827 for more details
Cross-reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK0827 is reactive to AGER in Human samples
Validated Sample Types
cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).
Application Guarantee
EK0827 is guaranteed for ELISA in Human by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of AGER
RAGE, the Receptor for Advanced Glycation Endproducts, is a 35kD transmembrane receptor of the immunoglobulin super family. It is also known as “AGER”. AGER gene is mapped to chromosome 6p21.3 by mapping by contiguous cosmids and YAC clones and by fluorescence in situ hybridization. The expression of RAGE is particularly increased in neurons close to deposits of amyloid beta peptide and to neurofibrillary tangles. RAGE has been linked to several chronic diseases, which are thought to result from vascular damage. The pathogenesis is hypothesized to include ligand binding upon which RAGE signals activation of the nuclear factor kappa B (NF-kappaB).
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK0827-CAP | Anti-Human AGER Pre-coated 96-well strip microplate | 1 |
| EK0827-ST | Human AGER Standard | 2 vials, 10 ng/tube |
| EK0827-DA | Human AGER 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 | 78 | 156 | 312 | 625 | 1250 | 2500 | 5000 |
| O.D. | 0.084 | 0.156 | 0.204 | 0.319 | 0.419 | 0.628 | 1.014 | 1.819 |
Data Example Images
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Human Rage PicoKine ELISA Kit standard curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect AGER, Dilution ratio of 1:1, concentration in serum and plasma is 294-5224 pg/ml..
Some articles we found to cite concentrations of AGER in samples: 20726839 (Pubmed IDs).
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Human Rage 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) | 154 | 623 | 3076 | 159 | 595 | 3344 |
| Standard deviation | 10.16 | 27.41 | 221.47 | 10.65 | 32.72 | 250.8 |
| CV (%) | 6.6% | 4.4% | 7.2% | 6.7% | 5.5% | 7.5% |
Specific Publications For Human Rage ELISA Kit PicoKine® (EK0827)
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Customer Reviews
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Customer Q&As
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7 Customer Q&As for Human Rage ELISA Kit PicoKine®
Question
Q: can you suggest the dilution ratio of serum samples for detection of AGER in Human serum? I am trying to measure a few 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.
P. Wilson
Verified customer
Asked: 2020-10-31
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 AGER 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: 2020-10-31
Question
Q: how long can samples (cell cultures, serum, and plasma) be stored and still be stable for analysis AGER using the EK0827 Human AGER Picokine® ELISA Kit?
Verified Customer
Verified customer
Asked: 2019-08-28
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 right after 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: 2019-08-28
Question
Q: how to proceed with the analysis of ELISA data? I measured AGER level in serum.
B. Johnson
Verified customer
Asked: 2019-01-26
Answer
A: please read 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: 2019-01-26
Question
Q: What is the optimal O.D. value for AGER ELISA kit? I performed your AGER 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 AGER even though the O.D. values are not very high?
Verified Customer
Verified customer
Asked: 2018-12-12
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. an important assessment 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 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 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 AGER ELISA kit to have sensitivity of 10pg/ml, that means the minimum amount of AGER that can be declared/interpreted as positive by the above standard is 10pg/ml.
Boster Scientific Support
Answered: 2018-12-12
Question
Q: what is the protocol regarding preparation of cell lysates prepared for use in Picokine® ELISA kits?
Verified Customer
Verified customer
Asked: 2018-08-31
Answer
A: for those Picokine® ELISAs where cell or tissue lysate is a validated sample type, sample preparation instructions for lysate are present 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: 2018-08-31
Question
Q: which procedure should I follow in order to thaw whole blood sample for AGER ELISA after freezing?
Z. Collins
Verified customer
Asked: 2018-08-03
Answer
A: we do not recommend freezing 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 AGER 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: 2018-08-03
Question
Q: Are Boster Bio recombinant proteins and antibodies sterile?
Verified Customer
Verified customer
Asked: 2017-05-31
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: 2017-05-31
