Mouse CD5L/CT-2 ELISA Kit PicoKine®

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.

Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 201 236 232 222 222 13.55 6.1% Sample 2 1789 1795 1875 1980 1859 77.28 4.1% Sample 3 4940 4445 4825 4834 4761 187.96 3.9% Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 201 236 232 222 222 13.55 6.1% Sample 1 201 236 232 222 222 13.55 6.1% Sample 2 1789 1795 1875 1980 1859 77.28 4.1% Sample 2 1789 1795 1875 1980 1859 77.28 4.1% Sample 3 4940 4445 4825 4834 4761 187.96 3.9% Sample 3 4940 4445 4825 4834 4761 187.96 3.9%

Specificaly expressed in tissue macrophages (PubMed:9892623) . Expressed in thymus, liver, spleen and lymph nodes (PubMed:10651944) . Present in Th17 cells; mainly present in non-pathogenic Th17 cells (PubMed:26607793) . .

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 201 1789 4940 207 1935 4821 Standard deviation 13.26 110.91 375.44 17.38 131.58 414.6 CV (%) 6.6% 6.2% 7.6% 8.4% 6.8% 8.6% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 201 1789 4940 207 1935 4821 Standard deviation 13.26 110.91 375.44 17.38 131.58 414.6 CV (%) 6.6% 6.2% 7.6% 8.4% 6.8% 8.6% Intra-Assay Precision Inter-Assay Precision Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 n 16 16 16 24 24 24 Mean (pg/ml) 201 1789 4940 207 1935 4821 Mean (pg/ml) 201 1789 4940 207 1935 4821 Standard deviation 13.26 110.91 375.44 17.38 131.58 414.6 Standard deviation 13.26 110.91 375.44 17.38 131.58 414.6 CV (%) 6.6% 6.2% 7.6% 8.4% 6.8% 8.6% CV (%) 6.6% 6.2% 7.6% 8.4% 6.8% 8.6%

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 (PubMed:26048980) . Able to inhibit lipid droplet size in adipocytes (PubMed:20519120, PubMed:22579686) . 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) (PubMed:20519120) . 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 (PubMed:21730133) . Regulation of intracellular lipids mediated by CD5L has a effect on transcription regulation mediated by nuclear receptors ROR-gamma (RORC) (PubMed:22579686, PubMed:26607793) . Acts as a key regulator of metabolic switch in T-helper Th17 cells (PubMed:26607794, PubMed:26607793) . 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 (PubMed:26607793) . 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 (PubMed:26607793) . 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 (PubMed:23562157) . Also acts as an inhibitor of apoptosis in macrophages: promotes macrophage survival from the apoptotic effects of oxidized lipids in case of atherosclerosis (PubMed:9892623, PubMed:16054063) . Involved in early response to microbial infection against various pathogens by acting as a pattern recognition receptor and by promoting autophagy (By similarity) . .

Description|Quantity Pre-coated 96-well strip microplate|1 Standard|2 vials Biotinylated antibody (100x) |100ul Avidin-Biotin-Peroxidase Complex (100x) |100ul Sample Diluent|30ml Antibody Diluent|12ml Avidin-Biotin-Peroxidase Diluent|12ml Color Developing Reagent (TMB) |10ml Stop Solution|10ml Wash Buffer (25x) |20ml Adhesive plate sealers|4

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Secreted . Cytoplasm . Secreted by macrophages and circulates in the blood (PubMed:20519120) . Transported in the cytoplasm via CD36-mediated endocytosis (PubMed:20519120) . .

Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.035 0.121 0.22 0.391 0.671 1.043 1.608 2.051 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.035 0.121 0.22 0.391 0.671 1.043 1.608 2.051 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.035 0.121 0.22 0.391 0.671 1.043 1.608 2.051 O.D. 0.035 0.121 0.22 0.391 0.671 1.043 1.608 2.051

Activate the sample Cell culture supernatants: add activating reagent pro rate, i.e. add 20μl of Solution A into 40μl of sample, 10 min later, add 20μl of Solution B. PH7.0-7.6. Plasma (heparin, EDTA), Serum: add activating reagent pro rate, i.e. add 20μl of Solution A into 40μl of sample, 10 min later, add 20μl of Solution B. PH7.0-7.6. Prior to the assay, the activated sample requires a 2000-fold dilution. Add 10μl of activated sample into 390μl wash buffer (1 : 40) . And then add 4μl of above diluted reagent into 196μl sample diluent buffer (1 : 50) . It is unnecessary to activate the recombinant CD5L. Sample was diluted partly after adding activating reagent, so please pay attention to this when calculate target protein concentration. Activate the sample Cell culture supernatants: add activating reagent pro rate, i.e. add 20μl of Solution A into 40μl of sample, 10 min later, add 20μl of Solution B. PH7.0-7.6. Plasma (heparin, EDTA), Serum: add activating reagent pro rate, i.e. add 20μl of Solution A into 40μl of sample, 10 min later, add 20μl of Solution B. PH7.0-7.6. Prior to the assay, the activated sample requires a 2000-fold dilution. Add 10μl of activated sample into 390μl wash buffer (1 : 40) . And then add 4μl of above diluted reagent into 196μl sample diluent buffer (1 : 50) . It is unnecessary to activate the recombinant CD5L. Sample was diluted partly after adding activating reagent, so please pay attention to this when calculate target protein concentration.

Microplate Reader capable of reading absorbance at 450nm. Incubator. Automated plate washer (optional) . Pipettes and pipette tips capable of precisely dispensing 0.5 μl through 1 ml volumes of aqueous solutions. Multichannel pipettes are recommended for large amount of samples. Deionized or distilled water. 500ml graduated cylinders. Test tubes for dilution.