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产品介绍
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Product Summary
The Quantikine Human LDL R Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human LDL R in cell culture supernates, cell lysates, serum, heparin plasma, and urine. It contains NS0-expressed recombinant human LDL R and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human LDL R showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human LDL R.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components
Cell Culture Supernates, Cell Lysates, Serum, Heparin Plasma, Urine
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 260 | 787 | 1598 | 287 | 823 | 1602 |
| Standard Deviation | 5.68 | 17.5 | 32.4 | 15.6 | 40 | 74.4 |
| CV% | 2.2 | 2.2 | 2 | 5.4 | 4.9 | 4.6 |
Recovery
The recovery of human LDL R spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 98 | 92-106 |
| Urine (n=4) | 94 | 91-99 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human LDL R were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay. Samples were diluted prior to assay.
Scientific Data
Assay Procedure
Refer to the product Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 50 µL of Assay Diluent to each well.
- Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
- Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
- Aspirate and wash 4 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
- Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
50 µL Assay Diluent
50 µL Standard, Control, or Sample
200 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Human LDLR Quantikine ELISA Kit Summary
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Cell Lysates (50 uL), Serum (10 uL), Heparin Plasma (10 uL), Urine (50 uL)
Sensitivity
10.3 pg/mL
Assay Range
62.5 - 4,000 pg/mL (Cell Culture Supernates, Cell Lysates, Serum, Heparin Plasma, Urine)
Specificity
Natural and recombinant human LDL R. This assay also recognizes recombinant human LDL R/PCSK9 complex.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
背景
别名
FH,FHC,LDL R,LDL receptor,LDLCQ2,LDLR,low density lipoprotein receptor,low-density lipoprotein receptor class A domain-containing protein 3,low-density lipoprotein receptor,Low Density Lipoprotein Receptor
背景
Background: LDLR
LDL R (Low Density Lipoprotein Receptor) is a widely expressed cell surface scavenger receptor. LDL R binds ApoB and ApoE, the apolipoproteins of low- and very low-density lipoproteins (LDL and VLDL), respectively. Hepatocyte LDL R is responsible for endocytosis and clearing of most plasma LDL cholesterol. At the low pH of the endocytic vesicle, it dissociates, allowing degradation of LDL and recycling of LDL R to the cell surface. Lack of LDL R expression or function causes familial hypercholesterolemia (FH). The protease PCSK9 can also cause increased plasma cholesterol by promoting LDL R degradation rather than recycling to the cell surface.
Long Name:
Low Density Lipoprotein Receptor
Entrez Gene IDs:
3949 (Human); 16835 (Mouse); 300438 (Rat); 396801 (Porcine); 102127361 (Cynomolgus Monkey)
Alternate Names:
FH; FHC; LDL R; LDL receptor; LDLCQ2; LDLR; low density lipoprotein receptor; low-density lipoprotein receptor class A domain-containing protein 3; low-density lipoprotein receptor
研究领域
制备和贮存
保存方式
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
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