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Mouse CD5L/CT-2 EZ-Set™ ELISA Kit (DIY Antibody Pairs)

Mouse CD5L/CT-2 ELISA Kit EZ-Set™ (DIY Antibody Pairs) (antibody pairs and standards for assay development) . Quantitate Mouse Cd5l in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA) .

Product Specifications

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.

Synonyms

CD5 antigen-like; Apoptosis inhibitor expressed by macrophages ; mAIM ; Apoptosis inhibitory 6; SP-alpha ; Cd5l; Aim , Api6

Gene Name

Cd5l

Gene ID

11801

UniProt

Q9QWK4

Reactivity

Mouse

Cross Reactivity

There is no detectable cross-reactivity with other relevant proteins.

Immunogen

Expression system for standard: NS0; Immunogen sequence: E22-V352

Tissue Specificity

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) . .

Applications

ELISA

Field of Research

Apoptosis, Cell Biology, Intracellular

Sample Type

Cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA) .

Detection Range

156 pg/mL - 10,000 pg/mL

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 (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) . .

Components

Catalog number Description Quantity Storage of opened/reconstituted material EZ1414-CA Goat anti-mouse CD5L/CT-2 polyclonal antibody (Capture Antibody) 500 μl, 4 μg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1414-DA Biotinylated goat anti-mouse CD5L/CT-2 polyclonal antibody (Detection Antibody) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EZ1414-ST Lyophilized recombinant mouse CD5L/CT-2 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material EZ1414-CA Goat anti-mouse CD5L/CT-2 polyclonal antibody (Capture Antibody) 500 μl, 4 μg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1414-DA Biotinylated goat anti-mouse CD5L/CT-2 polyclonal antibody (Detection Antibody) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EZ1414-ST Lyophilized recombinant mouse CD5L/CT-2 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material Catalog number Description Quantity Storage of opened/reconstituted material EZ1414-CA Goat anti-mouse CD5L/CT-2 polyclonal antibody (Capture Antibody) 500 μl, 4 μg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1414-CA Goat anti-mouse CD5L/CT-2 polyclonal antibody (Capture Antibody) 500 μl, 4 μg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1414-DA Biotinylated goat anti-mouse CD5L/CT-2 polyclonal antibody (Detection Antibody) 500 μl (recommended dilution 1:100) EZ1414-DA Biotinylated goat anti-mouse CD5L/CT-2 polyclonal antibody (Detection Antibody) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EZ1414-ST Lyophilized recombinant mouse CD5L/CT-2 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. EZ1414-ST Lyophilized recombinant mouse CD5L/CT-2 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit.

Storage Conditions

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.)

Calculated Molecular Weight

38863 MW

Specificity

Natural and recombinant mouse CD5L

Gene Name Synonym

CD5 antigen-like

Subcellular Location

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

Sequence Similarities

Contains 3 SRCR domains.

Protein Name

CD5 antigen-like

Sample Data

Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10,000 O.D. 0.030 0.115 0.210 0.345 0.557 0.900 1.384 1.909 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10,000 O.D. 0.030 0.115 0.210 0.345 0.557 0.900 1.384 1.909 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10,000 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10,000 O.D. 0.030 0.115 0.210 0.345 0.557 0.900 1.384 1.909 O.D. 0.030 0.115 0.210 0.345 0.557 0.900 1.384 1.909

Material Required But Not Provided

1. Microplate reader in standard size. 2. Automated plate washer. 3. Incubator. 4. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. 5. Clean tubes and Eppendorf tubes. 6. 96 well microplate (Cat# AR1100) 7. Plate Sealers. 8. Capture Antibody Diluent: PBS. 9. Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 μm filtered. 10. Substrate Solution: Tetramethylbenzidine (Cat# AR1104) 11. Stop Solution: 2 N H2SO4 (Cat# AR1105) 12. Wash Buffer (PBS and PBS-T) . Recipes: PBS: 8g NaCl, 0.2g KCl, 1.15g Na2HPO4, 0.2g KH2PO4, adjust the total volume to 1 L with distilled water, pH 7.2-7.4, 0.2 μm filtered. PBS-T: 0.1% Tween® 20 in PBS, pH 7.2-7.4. *Item 6 – 12 are included in the EZ Set Accessory Kit (EZA001)
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