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- ELISA Kit
- 人载脂蛋白B100(Apo-B100)ELISA Kit
This gene product is the main apolipoprotein of chylomicrons and low density lipoproteins. It occurs in plasma as two main isoforms, apoB-48 and apoB-100: the former is synthesized exclusively in the gut and the latter in the liver. The intestinal and the hepatic forms of apoB are encoded by a single gene from a single, very long mRNA. The two isoforms share a common N-terminal sequence. The shorter apoB-48 protein is produced after RNA editing of the apoB-100 transcript at residue 2180 (CAA->UAA), resulting in the creation of a stop codon, and early translation termination. Mutations in this gene or its regulatory region cause hypobetalipoproteinemia, normotriglyceridemic hypobetalipoproteinemia, and hypercholesterolemia due to ligand-defective apoB, diseases affecting plasma cholesterol and apoB levels.[provided by RefSeq]
- UniProtP04114
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BIOWWWAPOB
- HGNC603
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RGD2129
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MGI88052
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OMIM107730
- CUSABIOCSB-E08100h
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产品类型:
ELISA Kit
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产品名称:
人载脂蛋白B100(Apo-B100)ELISA Kit
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英文名称:
Human apolipoprotein B100 (Apo-B100) ELISA Kit
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货号:
CSB-E08100h
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别名:
FLDB, LDLCQ4, apoB-100|apoB-48|apolipoprotein B|apolipoprotein B48
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规格:
96T
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中文价格:
3600
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种属:
Homo sapiens (Human)
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其他种属:
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待测物名称:
apolipoprotein B (including Ag(x) antigen)
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缩写:
APOB
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蛋白功能1:
Apolipoprotein
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蛋白功能2:
Lipogenesis and lipometabolism
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蛋白功能3:
Cholesterol metabolism
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检测范围:
78 ng/mL-5000 ng/mL
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灵敏度:
19.5 ng/mL
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反应时间:
1-5h
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所需样本体积:
50-100ul
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检测波长:
450 nm
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下载:
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标准曲线:
| These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed. |
| ng/ml |
OD1 |
OD2 |
Average |
|
|
| 5000 |
0.074 |
0.073 |
0.074 |
|
|
| 2500 |
0.118 |
0.121 |
0.120 |
|
|
| 1250 |
0.182 |
0.172 |
0.177 |
|
|
| 625 |
0.272 |
0.284 |
0.278 |
|
|
| 312 |
0.451 |
0.442 |
0.447 |
|
|
| 156 |
0.770 |
0.754 |
0.762 |
|
|
| 78 |
1.122 |
1.054 |
1.088 |
|
|
| 0 |
2.371 |
2.270 |
2.321 |
|
|
|
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精密度:
| Intra-assay Precision (Precision within an assay): CV%<8% |
|
|
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| Three samples of known concentration were tested twenty times on one plate to assess. |
|
| Inter-assay Precision (Precision between assays): CV%<10% |
|
|
|
| Three samples of known concentration were tested in twenty assays to assess. |
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线性度:
| To assess the linearity of the assay, samples were spiked with high concentrations of human Apo-B100 in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay. |
| |
Sample |
Serum(n=4) |
|
| 1:200 |
Average % |
96 |
|
| Range % |
92-102 |
|
| 1:400 |
Average % |
95 |
|
| Range % |
91-99 |
|
| 1:800 |
Average % |
104 |
|
| Range % |
102-108 |
|
| 1:1600 |
Average % |
87 |
|
| Range % |
81-92 |
|
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回收率:
| The recovery of human Apo-B100 spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section. |
| Sample Type |
Average % Recovery |
Range |
|
| Serum (n=5) |
95 |
92-102 |
|
| EDTA plasma (n=4) |
90 |
87-94 |
|
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定性/定量:
quantitative
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研究领域:
Cardiovascular
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特异性：
This assay has high sensitivity and excellent specificity for detection of Homo sapiens (Human) APOB. No significant cross-reactivity or interference between Homo sapiens (Human) APOB and analogues was observed.
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精密度：
Intra-assay Precision (Precision within an assay): CV%<8%
Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10%
Three samples of known concentration were tested in twenty assays to assess.
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样本搜集及储存：
Serum: Use a serum separator tube (SST) and allow samples to clot for two hours at room temperature or overnight at 4°C before centrifugation for 15 minutes at 1000 ×g. Remove serum and assay immediately or aliquot and store samples at -20°C or -80°C. Avoid repeated freeze-thaw cycles.
Plasma: Collect plasma using EDTA, or heparin as an anticoagulant. Centrifuge for 15 minutes at 1000 ×g at 2-8°C within 30 minutes of collection. Assay immediately or aliquot and store samples at -20°C or -80°C. Avoid repeated freeze-thaw cycles.
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检测步骤：
Bring all reagents and samples to room temperature before use. Centrifuge the sample again after thawing before the assay. It is recommended that all samples and standards be assayed in duplicate.
1. Prepare all reagents, working standards, and samples as directed in the previous sections.
2. Refer to the Assay Layout Sheet to determine the number of wells to be used and put any remaining wells and the desiccant back into the pouch and seal the ziploc, store unused wells at 4°C.
3. Add 100μl of standard and sample per well. Cover with the adhesive strip provided. Incubate for 2 hours at 37°C. A plate layout is provided to record standards and samples assayed.
4. Remove the liquid of each well, don't wash.
5. Add 100μl of Biotin-antibody (1x) to each well. Cover with a new adhesive strip. Incubate for 1 hour at 37°C. (Biotin-antibody (1x) may appear cloudy. Warm up to room temperature and mix gently until solution appears uniform.)
6. Aspirate each well and wash, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (200μl) using a squirt bottle, multi-channel pipette, manifold dispenser, or autowasher, and let it stand for 2 minutes, complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining wash Buffer by aspirating ordecanting. Invert the plate and blot it against clean paper towels.
7. Add 100μl of HRP-avidin (1x) to each well. Cover the microtiter plate with a new adhesive strip. Incubate for 1 hour at 37°C.
8. Repeat the aspiration/wash process for five times as in step 6.
9. Add 90μl of TMB Substrate to each well. Incubate for 15-30 minutes at 37°C. Protect from light.
10. Add 50μl of Stop Solution to each well, gently tap the plate to ensure thorough mixing.
11. Determine the optical density of each well within 5 minutes, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. Subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.
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结果计算：
Using the professional soft "Curve Expert 1.3" to make a standard curve is recommended, which can be downloaded from our web.
Average the duplicate readings for each standard and sample and subtract the average zero standard optical density.
Create a standard curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve-fit. As an alternative, construct a standard curve by plotting the mean absorbance for each standard on the x-axis against the concentration on the y-axis and draw a best fit curve through the points on the graph. The data may be linearized by plotting the log of the APOB concentrations versus the log of the O.D. and the best fit line can be determined by regression analysis. This procedure will produce an adequate but less precise fit of the data.
If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.
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引用文献:
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1.
A study of serum Apo A-1 and Apo B-100 levels in women with preeclampsia ,
Timur H.et al,Pregnancy Hypertens.,6(2):121-5
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2.
Prevention mechanism of 2, 3, 5, 4'-tetrahydroxy-stilbene-2-O-β-D-glucoside on lipid accumulation in steatosis hepatic L-02 cell ,
Wang W.et al,J Ethnopharmacol. ,2017 May 14;153(3):763-70.
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3.
The barberry juice effects on metabolic factors and oxidative stress in patients with type 2 diabetes: A randomized clinical trial ,
FatemehLazavi.et al,Complementary Therapies in Clinical Practice,May
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