Human CD5L / CT 2 PicoKine™ Fast ELISA Kit


SKU FEK1413
Size 96wells/kit, with removable strips.
Reactivity Human
Sample Type cell culture supernates, cell lysates, serum and plasma (heparin, EDTA).
Sample Volume 100ul per well
Sensitivity <10pg/ml
Assay Range 156pg/ml-10000pg/ml

Introduction

Boster's Picokine™ Fast ELISA kits features the high sensitivity of Picokine™ ELISA kits and the significantly reduced assay run time. Now you can run the entire ELISA assay in less than 1.5 hours.

Compared to regular Picokine ELISA kits, the following sections in the datasheet are different:
1. kit components;
2. preparation before experiment;
3. assay protocol

Overview

Product Name Human CD5L / CT 2 PicoKine™ Fast ELISA Kit
SKU/Catalog Number FEK1413
Storage & Handling Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles(Shipped with wet ice.)
Size 96wells/kit, with removable strips.
Description The Fast version of Picokine ELISA kits, assay takes less than 1.5 hours. Detect Human CD5L with <10pg/ml sensitivity. Format: 96-well plate with removable strips. Compatible samples: cell culture supernates, cell lysates, serum and plasma (heparin, EDTA). This is a TMB colorimetric sandwich ELISA kit with short assay time and fast experiment set up. CD5L tissue specificity: Expressed in spleen, lymph node, thymus, bone marrow, and fetal liver, but not in non-lymphoid tissues. .
Cite This Product Human CD5L / CT 2 PicoKine™ Fast ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # FEK1413)
Sample Type cell culture supernates, cell lysates, serum and plasma (heparin, EDTA). Anticoagulant(s): heparin or EDTA
*The recommended anticoagulants are proven to not block the antibody binding sites on the target antigen. Please do not collect blood sample with other anticoagulants that are not specified above or contact us to check for feasibility.
Sensitivity <10pg/ml
Assay Range 156pg/ml-10000pg/ml
Immunogen Expression system for standard: NSO; Immunogen sequence: S20-G347
Reactivity Human
Cross Reactivity There is no detectable cross-reactivity with other relevant proteins.
Antibody Clonalities Capture Antibody | Detection Antibody:
monoclonal antibody from mouse|polyclonal antibody from goat

Assay Details

Kit Components

Catalog number Description Quantity
FEK1413-CAP 96-well plate precoated with anti-Human CD5L antibody 1
FEK1413-ST lyophilized recombinant Human CD5L standard 10ng/tube
FEK1413-DA biotinylated anti-Human CD5L antibody 130ul
AR1103 Avidin-Biotin-Peroxidase Complex(ABC) 130ul
AR1106-1 sample diluent buffer 30ml
AR1106-2 antibody diluent buffer 12ml
AR1106-3 ABC diluent buffer 12ml
AR1104 TMB color developing agent 10ml
AR1105 TMB stop solution 10ml
PLA-SEA Adhesive cover 4

*Why there is no wash buffer? Our Avidin-Biotin-Peroxidase Diluent contains the detergent (TWEEN) normally present in other companies' ELISA kits. This saves you the step of having to wash with the special wash buffer and achieve similar or better signal to noise ratio. The wash can use regular wash buffers (PBS, TBS etc.) commonly found in labs.

Materials Required But Not Provided

  • Microplate reader in standard size.
  • Automated plate washer.
  • Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection.
  • Clean tubes and Eppendorf tubes.
  • Washing buffer (neutral PBS or TBS).
  • Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g NaCl; 450μl of purified acetic acid or 700μl of concentrated hydrochloric acid to 1000ml H2
  • Preparation of 0.01 M PBS: Add 8.5g sodium chloride, 1.4g Na O and adjust pH to 7.2-7.

Typical Data Obtained from Human CD5L / CT 2 PicoKine™ Fast ELISA Kit

(TMB reaction incubate at 37°C for 15-20min)

Concentration(pg/ml)015631262512502500500010000
O.D.0.0080.4330.6771.0051.4091.6501.8492.099

Intra/Inter Assay Precision

Intra-Assay PrecisionInter-Assay Precision
Sample123123
n161616242424
Mean(pg/ml)2451172545022412885192
Standard deviation10.0479.69414.213.2191.44415.36
CV(%)4.1%6.8%7.6%5.9%7.1%8%

Reproducibility

Three samples with differing target protein concentrations were assayed using four different lots to measure the CV% lot to lot variance.

Reproducibility

To assay reproducibility, three samples with differing target protein concentrations were assayed using four different lots.

LotsLot1 (pg/ml)Lot2 (pg/ml)Lot3 (pg/ml)Lot4 (pg/ml)Mean (pg/ml)Standard DeviationCV (%)
Sample 12452422502442452.941.2%
Sample 21172125411481193119139.33.2%
Sample 354505193499454195264184.773.5%
*number of samples for each test n=16.

*The typical data is obtained from Boster's internal QC result and for reference only. It may differ from the lab test results of the end users. It is more important that the user's lab test results reflect the same linearity demonstrated in the typical data than achieving exactly the same O.D. values.

Target Info

Protein Target Info (Source: Uniprot.org)

Uniprot Id O43866
Gene Name CD5L
Protein Name CD5 antigen-like
Tissue Specificity Expressed in spleen, lymph node, thymus, bone marrow, and fetal liver, but not in non-lymphoid tissues. .
Alternative Names CD5 antigen-like;Apoptosis inhibitor expressed by macrophages ;hAIM ;CT-2 ;IgM-associated peptide ;SP-alpha ;CD5L;API6;UNQ203/PRO229 ;
Subcellular Localization Secreted . Cytoplasm . Secreted by macrophages and circulates in the blood (PubMed:24223991, PubMed:24804991). Transported in the cytoplasm via CD36-mediated endocytosis (By similarity). .
Molecular Weight 38088 MW

*if product is indicated to react with multiple species, protein info is based on the human gene.

Ontology

Protein Function Secreted protein that acts as a key regulator of lipid synthesis: mainly expressed by macrophages in lymphoid and inflammed tissues and regulates mechanisms in inflammatory responses, such as infection or atherosclerosis. Able to inhibit lipid droplet size in adipocytes. Following incorporation into mature adipocytes via CD36-mediated endocytosis, associates with cytosolic FASN, inhibiting fatty acid synthase activity and leading to lipolysis, the degradation of triacylglycerols into glycerol and free fatty acids (FFA). CD5L-induced lipolysis occurs with progression of obesity: participates to obesity-associated inflammation following recruitment of inflammatory macrophages into adipose tissues, a cause of insulin resistance and obesity- related metabolic disease. Regulation of intracellular lipids mediated by CD5L has a direct effect on transcription regulation mediated by nuclear receptors ROR-gamma (RORC). Acts as a key regulator of metabolic switch in T-helper Th17 cells. Regulates the expression of pro-inflammatory genes in Th17 cells by altering the lipid content and limiting synthesis of cholesterol ligand of RORC, the master transcription factor of Th17-cell differentiation. CD5L is mainly present in non-pathogenic Th17 cells, where it decreases the content of polyunsaturated fatty acyls (PUFA), affecting two metabolic proteins MSMO1 and CYP51A1, which synthesize ligands of RORC, limiting RORC activity and expression of pro-inflammatory genes. Participates in obesity- associated autoimmunity via its association with IgM, interfering with the binding of IgM to Fcalpha/mu receptor and enhancing the development of long-lived plasma cells that produce high-affinity IgG autoantibodies (By similarity). Also acts as an inhibitor of apoptosis in macrophages: promotes macrophage survival from the apoptotic effects of oxidized lipids in case of atherosclerosis (PubMed:24295828). Involved in early response to microbial infection against various pathogens by acting as a pattern recognition receptor and by promoting autophagy (PubMed:16030018, PubMed:24223991, PubMed:24583716, PubMed:25713983). .
Research Areas Human

*You can search these to find other products in these research areas.
Background CD5 antigen-like, also known as Sp alpha and AIM, is a protein that in humans is encoded by the CD5L gene. It is mapped to 1q21-q23 by fluorescence in situ hybridization. It is found that Aim expression is induced in mouse macrophages in response to loading with highly oxidized low density lipoprotein (oxLDL), and that Aim is expressed in foam cells within atherosclerotic lesions. Both the expression of Aim in lesions and its induction by oxLDL require Lxr /Rxr heterodimers. Aim-null macrophages are highly susceptible to oxLDL-induced apoptosis in vitro and undergo accelerated apoptosis in atherosclerotic lesions in vivo. Double knockout of Aim and Ldlr reduce atherosclerotic lesions. Therefore, it is concluded that AIM expression protects macrophages from apoptosis within atherosclerotic lesions, promoting early lesion development.

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FEK1413
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