Mouse Apolipoprotein B(APOB) ELISA kit

Code CSB-EL001918MO
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

This Mouse APOB ELISA Kit was designed for the quantitative measurement of Mouse APOB protein in serum, plasma, tissue homogenates. It is a Competitive ELISA kit, its detection range is 0.156 μg/mL-10 μg/mL and the sensitivity is 0.039 μg/mL.

Target Name Apolipoprotein B
Alternative Names ApobApolipoprotein B-100 ELISA kit; Apo B-100) [Cleaved into: Apolipoprotein B-48 ELISA kit; Apo B-48)] ELISA kit
Abbreviation APOB
Uniprot No. E9Q414
Species Mus musculus (Mouse)
Sample Types serum, plasma, tissue homogenates
Detection Range 0.156 μg/mL-10 μg/mL
Sensitivity 0.039 μg/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Cardiovascular
Assay Principle quantitative
Measurement Competitive
Precision
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.    
             
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of mouse APOB 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)  
1:100 Average % 102  
Range % 97-108  
1:200 Average % 103  
Range % 97-106  
1:400 Average % 92  
Range % 88-97  
1:800 Average % 86  
Range % 82-91  
Recovery
The recovery of mouse APOB 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) 94 89-98  
EDTA plasma (n=4) 105 102-109  
             
             
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
μg/ml OD1 OD2 Average    
10 0.091 0.092 0.092    
5 0.136 0.132 0.134    
2.5 0.236 0.229 0.233    
1.25 0.360 0.368 0.364    
0.625 0.539 0.524 0.532    
0.312 0.877 0.927 0.902    
0.156 1.334 1.286 1.310    
0 2.562 2.747 2.655    
Materials provided
  • A 96-well Assay plate --The 96-well plate has been pre-coated with ApoB.
  • Two pieces of Standard (Freeze-dried)--Dilute the standard at dilution series, read the OD values, and then draw a standard curve.
  • HRP-conjugated ApoB antibody (100 x concentrate) (1 x 60μl) --Act as the detection antibody.
  • HRP-conjugate Diluent (1 x 10 ml) --Dilute the HRP-conjugated ApoB antibody solution.
  • Sample Diluent (2 x 20 ml) --Reconstitute the standard and dilute the sample to an appropriate concentration.
  • Wash Buffer (25x concentrate) (1 x 20 ml)--Wash away unbound or free substances.
  • TMB Substrate (1x 10 ml) --Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • Stop Solution (1 x 10ml) --Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells)
  • An Instruction manual
Materials not provided
  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm - 570 nm.
  • An incubator that can provide stable incubation conditions up to 37°C±5°C.
  • Centrifuge
  • Vortex
  • Squirt bottle, manifold dispenser, or automated microplate washer
  • Absorbent paper for blotting the microtiter plate
  • 50-300ul multi-channel micropipette
  • Pipette tips
  • Single-channel micropipette with different ranges
  • 100ml and 500ml graduated cylinders
  • Deionized or distilled water
  • Timer
  • Test tubes for dilution
ELISA Data Analysis Watch ELISA data processing video & download Curve Expert if needed
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

Customer Reviews and Q&A

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 Q&A
Q:

hello, I want to install the analysis software, I can not install it, my computer does not have the necessary windows version.
Do you have a solution?

A:
Thanks for your inquiry! Here is our guidance on elisa data analysis, there is the software to download. Pls kindly note "Curve expert 1.3" or "Curve expert 1.4" is not compatible with Macs.
If you need the software for Macs, you can download here:
http://download.cnet.com/CurveExpert-Professional/3000-2054_4-75332285.html
It is a paid software. If you are not willing to use it, you can make the standard curve by Excel or other softwares, or you can send us your data and we will process the data for you.
Q:

Per customers request, please provide the following information regarding Mouse Apolipoprotein B, APOB ELISA Kit (CSB-EL001918MO)
1)The active ingredient of the stop solution and its concentration ?
2)The type, concentration, and state (liquid/lyophilized) of the preservatives used ?
3) If the components are lyophilized, what is the concentration prior to reconstitution (i.e. mass of preservative used/ total mass of lyophilized vial contents)?
4)Expression host of the standard ?

A:
Thanks for your inquiry!
CSB-EL001918MO
Stop solution: 4N sulfuric acid;
Preservatives is liquid proclin 300(1:3000);
The standard in the kit is lyophilized powder. The concentration is 10ug/mL aftering being resuspended by sample diluent.
The standard is purified crude extract of native samples.
Pls let me know if you have any further questions. Thank you.

Target Background

Function
(From Uniprot)
Apolipoprotein B is a major protein constituent of chylomicrons (apo B-48), LDL (apo B-100) and VLDL (apo B-100). Apo B-100 functions as a recognition signal for the cellular binding and internalization of LDL particles by the apoB/E receptor.
Gene References into Functions
  1. Type 2 diabetic, hyperlipidemic LDLr(-/-)ApoB(100/100) mice show increased calcific aortic valve disease. PMID: 29539583
  2. This study demonstrates that prevention of renal apoB accumulation is a mechanism by which TGF-beta inhibition is nephroprotective. PMID: 28912302
  3. Atorvastatin therapy did not show cholesterol-independent effects on inflammation in atherosclerotic lesions in Ldlr(-/-)Apob(100/100) mice, whereas a cholesterol-lowering diet intervention was effective. PMID: 28457625
  4. A peptide fragment of apoB-100 altered the immune-dominant epitope of CD8+ T cells and reduced atherosclerosis. PMID: 28711866
  5. enhanced VLDL TG secretion in the absence of hepatocyte ABCA1 is due to altered intracellular trafficking of apolipoprotein B (apoB), resulting in augmented TG addition to nascent VLDL. PMID: 28694219
  6. We carried out our experiment in mice deficient in the low density lipoprotein (LDL) receptor and expressing only ApoB100 molecule (ApoB - LDLr) where the development of atherosclerosis is known to closely mimic human atherosclerosis PMID: 27133569
  7. The effect of hypercholesterolemia induced immune response and inflammation on progression of atherosclerosis in ApoB(tm25gy) LDLr(tm1Her) mice, expressing only ApoB100 and deficient in the low density lipoprotein receptor. PMID: 26178198
  8. ApoB-containing lipoproteins contribute to augmentation of AngII-induced abdominal aortic aneurysms in male mice. PMID: 26044581
  9. Immunization with human apolipoprotein B100 (ApoB) resulted in four-fold increased accumulation of effector T cells in ApoB-containing matrigel PMID: 24866382
  10. Mice that produce apoB100 in the RPE and liver secrete lipoproteins into Bruch's membrane, but not to the extent that distinct features of AMD develop PMID: 25316721
  11. Its overexpression induce memory and cognitive decline in older animals, which indicates the importance of balanced lipid metabolism. PMID: 23643485
  12. The aim of this study was to characterize the ocular morphology of low-density lipoprotein receptor-deficient apolipoprotein B-100-only mice, with IGF-II overexpression. PMID: 23922490
  13. Our data establish the role of APOB gene in severe gut dysmotility. PMID: 22897442
  14. Cardiac lipotoxicity may originate from direct inhibition of myocardial ApoB lipoprotein and subsequent decreased lipid export, caused by supraphysiological levels of catecholamines. PMID: 23099354
  15. ApoB-100 transgenic animals present i) elevated serum and cerebral levels of triglycerides and ApoB-100, ii) increased cerebral tau phosphorylation at phosphosites Ser(199), Ser(199/202), Ser(396) and Ser(404). PMID: 23029362
  16. model in which increased hepatic sortilin binds intracellular APOB-containing particles in the Golgi apparatus as well as extracellular LDL at the plasma membrane and traffics them to the lysosome for degradation PMID: 22751103
  17. Hydrogen-saturated saline decreases athero-susceptibility in apoB-containing lipoprotein and aortic atherosclerosis in apoE-/- mice and improves HDL functionality in C57BL/6J mice. PMID: 22209213
  18. Echium oil is anti-atherogenic in atherosclerotic apoB100-only LDLrKO mice. PMID: 22100249
  19. knockdown of ApoB led to a significant reduction in serum lipids which was associated with an increase in hepatic triglycerides leading to liver steatosis in a mouse model having a human-like lipid profile PMID: 21826528
  20. Data suggest that ApoE associated with ApoB-carrying lipoproteins has an upregulatory role on ABCA1 expression, and that induction of Sp1 phosphorylation is a mechanism by which ApoE upregulates ABCA1 expression. PMID: 21779326
  21. Constitutive androstane receptor activation decreases plasma apolipoprotein B-containing lipoproteins and atherosclerosis in low-density lipoprotein receptor-deficient mice. PMID: 21778422
  22. siRNA-induced liver ApoB knockdown lowers serum LDL-cholesterol in a mouse model with human-like serum lipids. PMID: 21398511
  23. ApoB and HSP60 epitope immunization significantly reduces early atherosclerotic lesion in Apobtm2SgyLdlrtm1Her/J mice PMID: 20609438
  24. The transport of apolipoprotein (apo) B-100 in nonobese, nondiabetic, nonnephrotic CKD subjects and healthy controls (HC). PMID: 19542564
  25. Hypobetalipoproteinemic mice with a targeted apolipoprotein (Apo) B-27.6-specifying mutation: in vivo evidence for an important role of amino acids 1254-1744 of ApoB in lipid transport and metabolism of the apoB-containing lipoprotein. PMID: 11839763
  26. MTP is required for triacylglycerol (TG) accumulation in the microsomal lumen and as a source of TG for assembly with apoB, but normal levels of MTP are not required for transferring the bulk of TG to apoB during VLDL assembly in murine hepatocytes PMID: 12072432
  27. ApoB-48 and apoB-100 differentially influence the expression of type-III hyperlipoproteinemia in APOE*2 mice PMID: 12235184
  28. the developmental regulation of apoB mRNA editing is an autonomous cytodifferentiation function of small intestine and involves Cdx1 PMID: 12493769
  29. Deleting the C-terminal 20% from apoB-48 does not impair its ability to transport cholesterol and to support atherosclerosis, thus narrowing the "atherogenic region" of apoB. PMID: 12615667
  30. studies establish the existence of preferential degradation of intestinal apolipoprotein B-100 and subtle defects in triglyceride secretion in apolipoprotein B editing complex 1-/- mice PMID: 12816761
  31. secretion of apoB-100 from liver is impaired in hypobetalipoproteinemic mice with an apoB-38.9 allele PMID: 13130124
  32. cardiac apoB expression improves cardiomyopathy by increasing lipid resecretion from the heart PMID: 14634011
  33. transgenic hepatic lipase clears plasma cholesterol in LDL receptor-deficient "apoB-48-only" and "apoB-100-only" mice PMID: 14679168
  34. apolipoprotein B secretion is not dependent on dietary choline in mice PMID: 15024002
  35. Lipid peroxidation and oxidative stress regulate hepatic Apopb degradation and VLDL production. PMID: 15124019
  36. Data suggest the presence of additional regulatory regions outside the duplicated region of the apolipoprotein B gene, and that regulation of its expression is more complicated than previously thought. PMID: 15904874
  37. absence of expression of intestinal Apob48, but not Apob100, reduces biliary cholesterol secretion and cholelithogenesis, possibly by decreasing intestinal absorption and hepatic bioavailability PMID: 16175613
  38. In contrast, apoB38.9 mice effectively maintain cholesterol homeostasis in the liver, at least in part, by reducing hepatic cholesterol synthesis. PMID: 16455790
  39. ApoB lipoproteins that contain apoCIII increase THP-1 cell adhesion to ECs via PKCalpha and RhoA-mediated beta1-integrin activation. PMID: 16461842
  40. Together, results suggest in vivo and in vitro that wortmannin-sensitive PI3-kinases maintain a basal level of VLDL suppression PMID: 16798720
  41. CuZn-SOD deficiency causes ApoB degradation and induces hepatic lipid accumulation by impaired lipoprotein secretion in mice PMID: 16921198
  42. ApoB48, not ApoB100, is the preferred protein for the gut to coat chylomicrons to ensure efficient chylomicron formation and lipid absorption. PMID: 18006607
  43. These findings indicate that the majority of triglyceride destined for secretion by liver is synthesized by DGAT2. PMID: 18252207
  44. the LDLR directs apoB to degradation in a post-ER compartment; the reuptake mechanism of degradation occurs via internalization of apoB through a constitutive endocytic pathway and apoE through a ligand-dependent pathway PMID: 18272520
  45. These findings demonstrate that Ad-mMTP increases murine hepatic VLDL-TG secretion only in the apoB100 background, and even then only in situations with either increased hepatic TG accumulation or increased apoB100 expression. PMID: 18519977
  46. Rerport absence of hyperlipidemia/atherosclerosis in LDL receptor-deficient mice having apolipoprotein B100 without the putative receptor-binding sequences. PMID: 18617647
  47. apoB secretion in obese mice is regulated by arachidonate 5-lipoxygenase PMID: 18645210
  48. apoB100-LDL is a novel signaling molecule from the liver to peripheral fat deposits that may be an important link between atherogenic dyslipidemias and facets of the metabolic syndrome PMID: 19020660
  49. Perturbations in PTEN not only may influence hepatic insulin signaling and hepatic lipogenesis, but also may alter hepatic apoB-lipoprotein production and the MTP stability. PMID: 19026012
  50. Myocardial apoB may be an important cardioprotective system in settings such as myocardial ischemia and heart failure. PMID: 19460358

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Subcellular Location Cytoplasm. Secreted. Lipid droplet.
Database Links

KEGG: mmu:238055

STRING: 10090.ENSMUSP00000036044

UniGene: Mm.221239

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