Apolipoprotein Assays Kits: ApoA1, ApoB, ApoC3, ApoE

 

Homogeneous, cell-based quantification of human apolipopoproteins

The HTRF® apolipoprotein assays provide all necessary reagents for rapid and  sensitive quantification of human apolipoprotein A1, B, C3 or E in a homogeneous format. The assays may be used with cell culture supernatants, whole cells, lipoproteins and plasma. The assays are easily adaptable for 96- to 384-w formats and enables higher throughput compared to ELISA .

Apolipoproteins are constituents of triglyceride-rich lipoprotein including VLDL (or bad cholesterol) and HDL (or good cholesterol). They play a central role in regulating triglyceride metabolism and are key markers of cardiovascular diseases and  type 2 diabetes.

Stachel sequence-derived peptides to dissect aGPCR signaling pathways - Read scientific paper

Assay principle

The HTRF Apolipoprotein assays are sandwich immunoassays involving two monoclonal antibodies, one labelled with Eu-Cryptate (Donor) and the second with d2  (Acceptor). The intensity of the signal is proportional to the concentration of apolipoprotein present in the sample.

All four apolipoprotein assays are based on the same assay protocol and can be run with cell supernatants or whole cells, lipoproteins and human plasma. The antibodies labelled with HTRF fluorophores may be pre-mixed and added in a single dispensing step, to further streamline the protocol. The assay can be run in 96- to 384-well plates by simply resizing each addition volume proportionally. The figure on the right shows the Apo A1 assay protocol in a 384-well small volume white plate (20 µL final). When using the cell-based procedure, the detecting reagents are directly added to the assay plate following the stimultion step.

Product specifications

  Human Apo A1 Human Apo B Human Apo CIII Human Apo E
Incubation 2h at RT 3h at RT 2h at RT (or ON) 3h at RT (or ON)
Assay range 0 to 4000 ng/mL 0 to 4000 ng/mL 8 to 125 ng/mL 0.5 to 32 ng/mL
Detection limit 0.5 ng/mL 1.8 ng/mL 2.0 ng/mL 0.9 ng/mL
Extended range Up to 8,000 ng/ml Up to 8,000 ng/ml NA Up to 100 ng/ml
Cross reactivity

Low cross reactivity with: ApoB-100, ApoA2,

ApoC1, ApoC2, ApoC3, ApoE

Low cross reactivity with: ApoA1, ApoA2,

ApoC1, ApoC2, ApoC3, ApoE

Crossreactivity with ApoB-48: 100%

NA NA

(*) Based on 384-well low volume plate (20µL). Extending incubation period improves sensitivity

Cross-reactivity

Cross-reactivity (%) ApoA1 ApoB
ApoA1 100 0.004
ApoB-100 0.044 100
ApoA2 0.037 <0.001
ApoC1 0.014 <0.001
ApoC2 0.004 0.001
ApoC3 0.005 0.006
ApoE 0.004 0.004
ApoB-48 ND 100

Assay in action

Cell-based apolipoprotein assays to monitor Retinoid X receptor (RXR) agonist

Human hepatoma cells (HepG2) in 80-90% confluence were grown in DMEM (Gibco; 11885, low glucose) supplemented with 2% FBS. 5 μL of HepG2 cells (2,000 & 5,000 per well) and 5 μL of the RXR ligand 9-cis retinoic acid (9-cis-RA) in DMEM,  2% FBS at various concentrations were dispensed into a 384-well low volume black culture plate. After a 48 hour incubation at 37°C in CO2 atmosphere, 10 μL of the two pre-mixed HTRF® detection reagents (from either the ApoA1 or the ApoB assay kit) were added. Readout was performed on RUBYstar after 3 hours of incubation at room temperature (an extract from the poster by K Adachi).

Part#, inserts & MSDS

Ordering Info

DescriptionCat. noProduct insertMSDS
Human Apolipoprotein C3 kit - 500 tests63ADK001PEG
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Human Apolipoprotein C3 kit - 10,000 tests63ADK001PEH
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Human Apolipoprotein E kit - 500 tests63ADK004PEG
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Human Apolipoprotein E kit - 10,000 tests63ADK004PEH
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Human Apolipoprotein A1 kit - 500 tests64APAPEG
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Human Apolipoprotein A1 kit - 10,000 tests64APAPEH
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Human Apolipoprotein B kit - 500 tests64APBPEG
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Human Apolipoprotein B kit - 10,000 tests64APBPEH
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Companion products

DescriptionCat. noProduct insertMSDS
Diluent 1 - 20 ml62DL1DDD
Human Apolipoprotein C3 kit - Standard63ADK001CDA
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Human Apolipoprotein E kit - Standard63ADK004CDA
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