| Field | Specification |
|---|---|
| Mfr No | |
| Alternative Names | CD5 antigen-like;Apoptosis inhibitor expressed by macrophages ;hAIM ;CT-2 ;IgM-associated peptide ;SP-alpha ;CD5L;API6;UNQ203/PRO229 ; |
| Assay Time | |
| Assay Type | |
| Detection Range | |
| Expression System | |
| Gene ID | |
| Immunogen | Expression system for standard: NS0; Immunogen sequence: S20-G347 |
| Product Type | |
| Reactivity | |
| Sample Type(s) | cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). |
| Sensitivity | |
| Storage | |
| Target | |
| UniProt # |
Background
Also known as: CD5 antigen-like, Apoptosis inhibitor expressed by macrophages, hAIM, CT-2, IgM-associated peptide, SP-alpha, CD5L, API6.
Human CD5L/CT-2 (CD5L) is widely studied as a molecular readout in experimental models where changes in protein abundance reflect underlying biology. This target is frequently investigated in Molecular & Cellular Biology research contexts. This analyte is often discussed in the context of cell-surface signaling and cell-state markers. Many receptors and surface markers act as gateways for signaling or as phenotypic indicators of specific cell populations and activation states.
Biological context
In experimental systems, protein abundance can reflect regulated expression, secretion, processing, or clearance. Interpreting changes benefits from considering compartment (cell-associated vs soluble), the time scale of regulation, and whether complexes or modified forms contribute to the measured signal.
Why it matters in research
- Systems-level readout: Quantification supports comparisons across conditions, time points, and treatment groups.
- Mechanistic interpretation: Pairing with upstream regulators and downstream markers helps contextualize changes.
- Biomarker-style profiling: Measuring panels of related analytes can improve interpretability in complex models.
Sample data
| Concentration (pg/ml) | 0 | 156 | 312 | 625 | 1250 | 2500 | 5000 | 10000 |
| O.D. | 0.031 | 0.094 | 0.157 | 0.282 | 0.507 | 1.032 | 1.901 | 2.669 |
Intra/inter assay consistency
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 16 | 16 | 16 | 24 | 24 | 24 |
| Mean (pg/ml) | 245 | 1172 | 5450 | 224 | 1288 | 5192 |
| Standard deviation | 10.04 | 79.69 | 414.2 | 13.21 | 91.44 | 415.36 |
| CV (%) | 4.1% | 6.8% | 7.6% | 5.9% | 7.1% | 8% |
Kit components
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|4Materials required but not provided
- 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.
Activating reagent preparation
CD5 combines with its ligand (CD5L) to form complex in samples. Free CD5L needs to be released by activation.
Solution A: 1N HCI: add 8.33ml of 12N HCI into 91.67ml of H2O.
Solution B: 1.2N NaOH/0.5M HEPES: add 12ml of 10N NaOH and 11.9g HEPES into 75ml of H2O, add H2O to adjust volume to 100ml.
Sample activation procedure
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.
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).
Plasma(heparin, EDTA): 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 4000-fold dilution. Add 10μl of activated sample into 790μl wash buffer (1 : 80). And then add 4μl of above diluted reagent into 196μl sample diluent buffer (1 : 50).Optimal dilutions should be determined by end users.
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.
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- Savaş et al. (2020). Apoptosis Inhibitor of Macrophage, Monocyte Chemotactic Protein-1, and C-Reactive Protein Levels Are Increased in Patien…. Metabolic Syndrome and Related Disorders.