Human Apolipoprotein A-I ELISpot Development Module, 5 Plate

Catalog #: SEL3664 Datasheet / COA / SDS

Discontinued Product

SEL3664 has been discontinued.
View all Apolipoprotein A-I/ApoA1 products.
R&D Systems ELISpot Development Modules
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Human Apolipoprotein A-I ELISpot Development Module, 5 Plate Summary

Assay Type
Quantitative Sandwich ELISA Development Module
Format
96-well microplate, sold separately (see Other Reagents Required)
Assay Length
3 hours 35 mins to 4 hours 50 mins**
Sample Type
Whole Cells
Sufficient Materials
For five 96-well microplates
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.


PRODUCT SUMMARY
Complete ELISpot kits are highly sensitive, microplate-based assays for the detection of cytokine secreting cells. Kits are available for detection and enumeration of a single analyte or two analytes simultaneously. Complete ELISpot kits are ready-to-run and require no assay development or refinement. ELISpot Development Modules contain the basic components required to develop an ELISpot assay. They offer an economical alternative to buying separate antibodies.

ELISpot development modules are an alternative to ELISpot kits. A basic understanding of ELISpot assay development is required for the successful use of these reagents. Each investigator should optimize the coating conditions, the assay sensitivity, the type of enzyme and substrate, as well as the concentrations of the capture and detection antibodies to achieve desired results. The analyte-specific ELISpot Development Module and the ELISpot Blue Color Module contain the necessary components for analyte detection and visualization, respectively. These modules can be used together but are sold separately. Each module contains enough reagents for at least five 96-well microplates.


PRODUCT FEATURES

  • An economical alternative to ELISpot Kits
  • Optimized capture and detection antibody pairings and recommended concentrations save lengthy development time
  • Generic development protocols provide direction to start an optimization protocol
  • Customize the assay to your specific needs


MODULE CONTENTS

  • Human Apolipoprotein A-I/ApoA1 Capture Antibody
  • Human Apolipoprotein A-I/ApoA1 Biotinylated-conjugated Detection Antibody


OTHER REAGENTS REQUIRED

  • ELISpot Blue Color Module or equivalent (R&D Systems, Catalog # SEL002)
  • PBS - 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
  • Wash Buffer - 0.05% Tween® 20 in PBS
  • Blocking Buffer - 1% BSA, 5% Sucrose in PBS
  • Reagent Diluent - 1% BSA in PBS, pH 7.2 - 7.4, 0.2 µm filtered
  • 2 °C – 8 °C refrigerator
  • 37 °C CO2 incubator
  • Positive Control - Use Recombinant Human Apolipoprotein A-I/ApoA1 or cells known to secrete Human Apolipoprotein A-I/ApoA1
  • 96-well plates - Nitrocellulose-bottom plates, PVDF-bottom Immunospot® plates, or flat-bottom polystyrene Immulon® ELISA plates
  • Multi-channel pipette, squirt bottle, manifold dispenser, or automated microplate washer
  • Dissection microscope or an automated ELISpot Reader
  • Deionized H2O

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Product Datasheets

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

Background: Apolipoprotein A-I/ApoA1

The apolipoproteins are a structurally-unrelated group of proteins that have some association with the transport of lipids in blood. Apolipoproteins, plus phospholipids, cholesterol and triglycerides, form spherical particles with a lipid/hydrophobic center and a (apolipo)protein coat. The apolipoprotein coat promotes aqueous solubility and serves as a ligand for lipoprotein receptors. HDL may contain apolipoproteins A, C, D, E, J, L and M, while LDL contains apolipoproteins B and E.

ApoAI and ApoA2 are major protein components of serum high-density lipoprotein (HDL) and are produced by the liver and small intestine. They are involved in reverse cholesterol transport from tissues to the liver. Polymorphisms of ApoA2 are associated with disorders of cholesterol and fatty acid metabolism. Human ApoB (Apolipoprotein B-100) is a 550 kDa, secreted, palmitoylated glycoprotein that is part of LDL and VLDL particles. It is made by liver and is 4536 aa in length. It binds LDL to the ApoB/E receptor. ApoC activates lipoprotein lipase and may self-associate to form amyloid-type fibrils.

ApoE is a 34 kDa protein component of serum chylomicrons, VLDL, and HDL particles. It mediates the binding, uptake, and catabolism of these particles through interactions with the ApoE receptor and LDL receptors in the liver and brain. ApoE is important in fatty acid homeostasis and memory formation. Polymorphisms encode three variants (ApoE2, 3, 4) which are differentially related to the development of atherosclerosis and neurogenerative disorders, particularly Alzheimer's disease.

Serum amyloid A proteins (SAAs) are a family of homologous apolipoproteins of high density lipoprotein (HDL). They can be divided into two groups. The first group consists of the acute phase SAA1 and SAA2 that associate with HDL during inflammation and remodel the HDL particle by displacing apolipoprotein A1. The second group consists of constitutively expressed SAA4 and SAA5 that exist as minor apolipoproteins on HDL but make up more than 90% of the total SAA during homeostasis.

Entrez Gene IDs:
335 (Human); 11806 (Mouse); 25081 (Rat)
Alternate Names:
Alp-1; APOA1; Apo-AI; apoA-I; Apolipoprotein A1; Apolipoprotein AI; Apolipoprotein A-I; Brp-14; Ltw-1; Lvtw-1; MGC117399; Sep-1; Sep-2

FAQs

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