Recombinant Human CD5L Protein

Cat. No.: rp144064

GRADE & PURITY Animal Free, Carrier Free, Bioactive, ActiBioPure™, High Performance, His Tag, ≥95%(SDS-PAGE)

Accession #

O43866

Protein Tag

C-His

Expression system

HEK293

Endotoxin Concentration

<0.1 EU/μg

Bioactivity

Measured by its ability to inhibit apoptosis on the J774A.1 mouse reticulum cell sarcoma macrophage cells. The presence of rhCD5L inhibited rhFas Ligand induced apoptosis by 70-90%. The ED50 for this effect is 1-4 µg/mL.

Vue d’ensemble

Function
May play a role in the regulation of the immune system. Seems to play a role as an inhibitor of apoptosis.

Specifications

Product Name

Recombinant Human CD5L Protein

Synonymes

Apoptosis inhibitor expressed by macrophages | CT-2 | hAIM | IgM-associated peptide | SP-alpha | API6 | AIM | apoptosis inhibitor 6 | CD5 antigen-like (scavenger receptor cysteine rich family) | CD5 antigen-like | CD5 molecule-like | CD5L | CT-2 | IgM-ass

Grade

ActiBioPure™, Animal Free, Bioactive, Carrier Free, High Performance, His Tag

Spécifications et pureté

Animal Free,Carrier Free,Bioactive,ActiBioPure™,High Performance,His-Tag,≥95%(SDS-PAGE)

Bioactivité

Measured by its ability to inhibit apoptosis on the J774A.1 mouse reticulum cell sarcoma macrophage cells. The presence of rhCD5L inhibited rhFas Ligand induced apoptosis by 70-90%. The ED50 for this effect is 1-4 µg/mL.

Concentration d'endotoxine

<0.1 EU/μg

Système d'expression

HEK293

Espèces

Human

Acides aminés

20-347 aa

Séquence

SPSGVRLVGGLHRCEGRVEVEQKGQWGTVCDDGWDIKDVAVLCRELGCGAASGTPSGILYEPPAEKEQKVLIQSVSCTGTEDTLAQCEQEEVYDCSHDEDAGASCENPESSFSPVPEGVRLADGPGHCKGRVEVKHQNQWYTVCQTGWSLRAAKVVCRQLGCGRAVLTQKRCNKHAYGRKPIWLSQMSCSGREATLQDCPSGPWGKNTCNHDEDTWVECEDPFDLRLVGGDNLCSGRLEVLHKGVWGSVCDDNWGEKEDQVVCKQLGCGKSLSPSFRDRKCYGPGVGRIWLDNVRCSGEEQSLEQCQHRFWGFHDCTHQEDVAVICSGHHHHHH

Journée des protéines

C-His

Accession #

O43866

Poids moléculaire prévu

36.9 kDa

SDS-PAGE

43 kDa, under reducing conditions

Type de molécule

Protein

Stockage et expédition

Forme

Lyophilized

Reconstitution

Reconstitute in sterile water to a concentration of 0.1-0.5 mg/ml.

Conditions de stockage de stockage

Store at -20°C,Avoid repeated freezing and thawing

Expédié en

Ice chest + Ice pads

Stabilité et stockage

Store at -20~-80℃ for more than 1 year. Upon delivery aliquot. Avoid freeze/thaw cycle.

Images

SDS-PAGE: Recombinant Human CD5L Protein (rp144064)

Recombinant Human CD5L Protein (rp144064) - SDS-PAGE
2 μg/lane of Recombinant Human CD5L Protein was resolved with SDS-PAGE under reducing (R) and non-reducing (N) conditions and visualized by Coomassie® Blue staining, showing the band at 42 kDa under reducing conditions and 30 kDa under non-reducing conditions.

Application Protocols

The following are general, literature-informed protocols for common uses of Carrier-Free, His-tagged recombinant proteins like CD5L. Adjust to your specific assay and consult the CoA/Spec Sheet for any item-specific instructions.

1. Reconstitution and aliquoting (general)

• Briefly centrifuge the vial. Add sterile, room-temperature buffer (e.g., PBS or HEPES, pH 7.2–7.6) to the desired concentration. Gently swirl—do not vortex.
• Optional: After initial dissolution, add 5–10% glycerol for storage; add 0.1% BSA only if adsorption losses are problematic (note: product is supplied Carrier Free).
• Filter-sterilize through 0.22 μm if sterility is critical. Aliquot, snap-freeze on dry ice/ethanol, and store at −20°C.

2. ELISA standard curve (antigen format)

• Coat capture antibody on high-binding plate per vendor instructions.
• Prepare serial 2–3-fold dilutions of CD5L (e.g., 1,000 to 1 ng/mL) in assay buffer with 0.1% protein and 0.05% Tween-20.
• Incubate, wash, and detect with validated detection antibody. Generate 4PL/5PL curve.

3. BLI/SPR binding assay

• Load His-tagged CD5L onto NTA/Ni sensors or capture via anti-His.
• Equilibrate in HBS-P (10 mM HEPES, 150 mM NaCl, 0.005–0.05% surfactant), pH 7.4.
• Inject analyte over 0.1–1,000 nM; fit association/dissociation to derive KD.

4. Cell treatment

• Prepare working stocks in serum-free medium; treat cells at 1–1,000 ng/mL.
• Include vehicle and protein controls; monitor endpoints (e.g., qPCR, flow cytometry, lipid uptake assays).

Biological Roles

The following summarizes general, literature-based biology of human CD5L (CD5 antigen-like; AIM/Api6). These points describe biochemical and cellular functions and do not imply clinical/therapeutic use.

• Family and structure: A secreted member of the scavenger receptor cysteine-rich (SRCR) protein family. The SRCR domains confer binding to diverse ligands, often via basic surfaces and disulfide-stabilized modules.
• Expression and source: Produced predominantly by tissue macrophages and certain immune cells (literature). Levels can vary with immune activation states.
• Molecular functions (in vitro/literature):
  • Binds specific lipids and modified lipoproteins, contributing to lipid handling by myeloid cells.
  • Acts as an opsonin-like factor in some contexts, modulating uptake/clearance pathways.
  • Interacts with select cell-surface receptors/coreceptors implicated in innate immune sensing and homeostasis.
• Pathway context: Associated with processes in innate immunity, apoptotic cell handling, and metabolic-immune crosstalk (literature). Exact partners and affinities depend on assay format, glycoform, and ionic conditions.
• Experimental implications: Recombinant CD5L can be used to probe macrophage polarization markers, lipid uptake/efflux readouts, and signaling pathway activation (e.g., NF-κB reporters) under defined conditions.

Note: Activity is highly context-dependent (cell type, differentiation state, co-factors such as calcium and lipids). Validate dose–response curves and include appropriate protein and buffer controls.

Buffer Applications

This protein is typically used in near-physiological aqueous buffers. Exact formulation for this item is not specified; refer to CoA/Spec Sheet. Below are general-purpose buffer systems and practical preparation notes.

• Phosphate-buffered saline (PBS), pH 7.2–7.6: 10 mM phosphate, 137 mM NaCl, 2.7 mM KCl. Suitable for most ELISAs, Westerns, and general handling.
• HEPES-buffered saline (HBS), pH 7.4–7.6: 10–20 mM HEPES, 150 mM NaCl. Better pH stability at ambient/CO2-independent conditions and in low-buffered media.
• Tris-buffered saline (TBS), pH 7.4–8.0: 20–50 mM Tris, 150 mM NaCl. Useful where phosphate interferes (e.g., certain metal chelation or calcium-dependent assays).
• Additives (as compatible with downstream assays):
  • 0.05–0.1% carrier protein (BSA) to mitigate adsorption losses in low-protein systems (note: product is supplied Carrier Free).
  • 5–10% glycerol for freeze–thaw protection; 0.001–0.02% Tween-20 to reduce surface binding.
  • Avoid EDTA or chelators when working with Ni-NTA capture of His-tag or when divalent cations are required for interactions.

Preparation tips:

• Filter sterilize (0.22 μm) and chill buffers before use.
• Titrate ionic strength (100–300 mM NaCl) to balance solubility vs. specific binding.
• Validate pH at working temperature; Tris pKa is temperature sensitive.

Green Alternatives

For recombinant proteins, the “greenness” considerations focus on buffer systems, preservatives, and cold-chain logistics rather than replacement chemicals.

• Prefer aqueous buffers over organic co-solvents. Avoid large volumes of acetonitrile/methanol except when required for MS sample prep.
• Use HEPES or phosphate buffers prepared from high-purity salts; avoid azide where possible (toxic to aquatic life), substituting sterile filtration and low-temperature storage for bioburden control.
• Optimize stability to reduce waste: aliquot to eliminate freeze–thaw loss; include small amounts of glycerol (5–10%) if compatible, allowing storage at −20°C without repeated cycles.
• Packaging and shipping: Consolidate orders to minimize cold-chain shipments. Reuse/recycle ice packs and insulating materials when allowed by institutional policy.

Illustrative comparison (general):

• Sodium azide vs sterile technique: Azide offers antimicrobial protection but adds toxicity and disposal burdens; sterile filtration (0.22 μm) and aseptic handling can maintain sterility without azide for short-term use.
• High-organic LC-MS buffers vs aqueous capture assays: Where feasible, use aqueous capture/detection (ELISA, BLI) rather than LC-MS requiring organic solvents, reducing solvent use.

Net effect: Careful buffer selection and handling practices can meaningfully reduce environmental impact without compromising protein performance.

Pharmaceutical Uses

This product is provided strictly for research use only (non-clinical, non-diagnostic). There are no claims of compliance with pharmacopeial monographs, and no authorization for human or veterinary administration.

Potential roles in pharmaceutical research workflows (general, non-clinical):

• Reference standard or antigen in immunoassay development (e.g., screening anti-CD5L antibodies).
• Tool protein for target validation, mechanism-of-action studies, or pathway interrogation in discovery biology.
• Reagent in high-content phenotypic screens assessing macrophage or lipid-handling biology under defined conditions.

Formulation/excipient status: Not applicable. The item is not supplied as an excipient and carries no therapeutic-use designation.

Regulatory note: For any use in regulated studies, obtain and review batch-specific documentation (CoA/Spec Sheet), including identity, purity, activity, and endotoxin specifications as required by your QA/QC procedures.

Physical Properties

Item-specific physicochemical specifications (appearance, buffer composition, exact MW, concentration) are not provided in the product data sheet.

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight (observed): Not specified for this item; refer to CoA/Spec Sheet.
  • Literature: CD5L monomer typically migrates ~38–40 kDa under reducing SDS-PAGE; glycosylation and tags can shift apparent size (literature).
• Isoelectric point (pI): Not specified for this item; literature pI values vary by glycoform and sequence construct.
• Extinction coefficient (ε280): Not specified for this item; can be computed from the exact sequence once provided (general guidance).
• Solubility: Recombinant proteins are generally soluble in physiological aqueous buffers (e.g., PBS, HEPES, Tris) with salt (100–300 mM NaCl) (general guidance). Optimize by gentle titration of pH (6.5–8.0), ionic strength, and optional additives (e.g., 5–10% glycerol) as compatible with assays.
• Formulation/refractive index/density/MP/BP/logP: Not applicable or not specified for this item.

Notes for practitioners:

• Minimize mechanical shear and foam; handle on ice.
• For concentration determinations post-reconstitution, use A280 with the construct-specific ε (once sequence is known) or a colorimetric protein assay compatible with your buffer (e.g., BCA avoiding reducing agents when not compatible).

Quality and Grades

This item is supplied with multiple performance designations. The following explains their typical meaning and practical implications for experimental design.

• Animal Free: Manufactured without animal-derived components to minimize risk of adventitious agents and batch-to-batch variability from animal sources. Suitable for workflows seeking xeno-free conditions.
• Carrier Free: No added carrier proteins (e.g., BSA, HSA) in the supplied formulation. Advantages: maximal specific activity and freedom to add your own carriers if needed. Consider adding a carrier (e.g., 0.1% BSA) only after initial reconstitution if adsorption losses are problematic in low-protein matrices (general guidance).
• Bioactive: Produced and qualified to retain functional activity in a relevant in vitro assay. Exact method and unit definition are not specified for this item; refer to CoA/Spec Sheet for the assay used.
• ActiBioPure™: Aladdin Scientific performance designation indicating high purity and maintained activity for bioassays. Specific purity metrics (e.g., SDS-PAGE %) are not specified; refer to CoA/Spec Sheet.
• High Performance: Denotes tight release criteria for purity/activity/identity suitable for demanding research applications. Item-specific thresholds not provided; refer to CoA/Spec Sheet.
• His Tag: Polyhistidine affinity tag enabling nickel/cobalt IMAC purification and facilitating detection/capture. Tag location (N- vs C-terminal) and linker composition are not specified for this item.

Batch-specific data (purity %, endotoxin level, host cell protein/DNA, buffer composition, concentration) are Not specified for this item; refer to CoA/Spec Sheet.

Reaction and Applications

While not a small-molecule reagent, recombinant CD5L is widely used as a functional protein standard and stimulus in biochemical and cell-based studies. Applications below are based on general literature for CD5L and related SRCR proteins; consult your protocol for specifics.

• Ligand–receptor studies: Assess binding to candidate receptors or co-receptors on myeloid cells using ELISA-type capture, biolayer interferometry (BLI), or SPR. His-tag enables oriented capture on NTA/Ni surfaces.
• Macromolecular interaction mapping: Identify CD5L interaction partners by pull-down from cell lysates using Ni-agarose or anti-His resin followed by MS-based proteomics (ensure mild lysis buffers to preserve interactions).
• Functional cell assays: Evaluate effects on macrophage/monocyte phenotypes, lipid handling, or cytokine responses by treating primary cells or lines with dose ranges (e.g., 1–1,000 ng/mL; optimize empirically). Avoid endotoxin artifacts by verifying endotoxin and including polymyxin B controls when appropriate.
• Standard/calibrator: Use as a positive control antigen in immunoassays (e.g., anti-CD5L ELISA) to generate standard curves and verify assay linearity.
• Structural/biophysical: Characterize oligomeric state by SEC-MALS or AUC; examine glycoform heterogeneity by deglycosylation and LC-MS.

Practical tips:

• Pre-clear matrices with Ni-NTA to reduce nonspecific His-mediated binding.
• Include matched-tag controls to distinguish tag-mediated vs native interactions.
• Confirm activity in your system since bioactivity can be context-dependent (glycoform, tag orientation, buffer, and temperature).

Reaction Conditions

For proteins, “reaction conditions” correspond to assay conditions. Exact activity assay used for this item is not specified; refer to CoA/Spec Sheet. The following are general conditions for common use-cases with His-tagged recombinant proteins.

• Binding assays (ELISA/BLI/SPR):
  • Buffer: PBS or HEPES-buffered saline, pH 7.2–7.6; 0.005–0.05% Tween-20 to minimize nonspecific binding.
  • Temperature: 20–25°C for BLI/SPR; 4–25°C for ELISA incubations.
  • Immobilization: Capture via anti-His antibody or Ni-NTA surfaces; avoid EDTA/strong chelators; include 150–300 mM NaCl to reduce nonspecific interactions.
  • Analyte ranges: 0.1–1,000 nM depending on affinity; perform serial dilutions spanning anticipated KD (general guidance).
• Cell-based treatments:
  • Medium: Serum-free or low-serum during treatment to minimize sequestration by serum proteins; reintroduce serum post-treatment as needed.
  • Dose: Start 1–1,000 ng/mL; optimize by monitoring phenotypic or transcriptional readouts.
  • Time: 1–48 h depending on endpoint (signaling vs transcriptional responses).
• Pull-down/CoIP:
  • Lysis: Mild, non-denaturing buffers (e.g., 20 mM HEPES, 150 mM NaCl, 0.5% NP-40/Triton X-100 equivalent), protease inhibitors; keep at 4°C.
  • Wash: Incremental salt (150–500 mM) and low detergent to reduce nonspecific binders.

Always benchmark conditions with positive/negative controls and verify protein integrity by SDS-PAGE/Western prior to and after assays.

Safety and Handling

• GHS classification: Not specified for this item; refer to SDS.
• Signal word / H-statements / pictograms: Not specified for this item; refer to SDS.

General laboratory safety for recombinant proteins (guidance; defer to SDS):

• PPE: Laboratory coat, gloves, and safety glasses. Avoid creating aerosols; work in a clean area or biological safety cabinet if sterility is required.
• Avoid repeated freeze–thaw cycles (can denature protein). Aliquot immediately after first thaw.
• Incompatibilities: Strong oxidants, extreme pH, proteases, and microbial contamination can degrade protein integrity.
• First aid (general):
  • Inhalation: Move to fresh air; seek medical attention if symptoms persist.
  • Skin/eye contact: Rinse with water for several minutes; remove contaminated clothing; seek medical attention if irritation persists.
  • Ingestion: Rinse mouth; do not induce vomiting; seek medical attention.
• Spill response: Absorb with inert material; clean with suitable disinfectant if sterility/biomaterial concerns. Dispose according to institutional guidelines for biological materials.
• Waste: Dispose as laboratory biological/chemical waste per local regulations.

Always consult the product-specific SDS and institutional biosafety procedures for authoritative guidance.

Solvent Selection

This product is a recombinant protein intended for use in aqueous buffers rather than organic solvents.

• Primary medium: Aqueous physiological buffers (e.g., PBS, HEPES, Tris; pH typically 7.2–7.8) with moderate ionic strength (100–300 mM NaCl) to promote solubility and stability (general guidance).
• Additives (as assay-compatible): 5–10% glycerol for freeze–thaw protection; 0.001–0.02% nonionic detergent (e.g., Tween-20) to reduce surface adsorption; 0.1% carrier protein (BSA) only if adsorption losses occur (note: item is supplied Carrier Free).
• Avoid: Strongly denaturing solvents (acetonitrile, methanol, DMSO) except for specialized applications (e.g., LC-MS sample prep); extremes of pH; high concentrations of chaotropes unless intentionally unfolding.

Comparison (general):

• PBS vs HEPES: PBS is convenient and inexpensive; HEPES maintains stable pH under CO2-independent conditions and at low buffer dilution.
• Tris: Good buffering near neutral pH; temperature-dependent pKa may affect pH on ice vs room temperature.

Tip: For cell-based assays, ensure buffer osmolality and pH match culture conditions. For binding assays, include appropriate divalent cations or chelators based on cofactor dependence of the interacting partner (if known).

Storage and Reconstitution

• Storage conditions (as provided): Store at −20°C. Avoid repeated freezing and thawing.
• Shipping: Ice chest with ice pads.

Item-specific formulation, concentration, and appearance: Not specified for this item; refer to CoA/Spec Sheet.

General reconstitution guidance for Carrier-Free, His-tagged recombinant proteins:

• Thaw the vial on ice. Briefly spin down contents.
• Reconstitute with sterile buffer (e.g., PBS or HEPES, pH 7.2–7.6) to a convenient stock concentration. Gently mix without vigorous agitation.
• Optional stabilizers (if assay-compatible): 5–10% glycerol to protect during freezing; add 0.1% carrier protein (e.g., BSA) only after initial dissolution if adsorption losses are an issue (note: supplied Carrier Free).
• Aliquot immediately into single-use volumes; snap-freeze and store at −20°C or below. Avoid frost-free freezers.

Handling tips:

• Keep on ice during use; minimize time at room temperature.
• Do not refreeze thawed aliquots; discard leftovers after use.
• For long-term storage or highly sensitive assays, consider storing at ≤−70°C if compatible with formulation (general guidance).

Always consult the lot-specific CoA/Spec Sheet for any special instructions (e.g., reconstitution volume, buffer composition, observed molecular weight, and bioactivity assay).

Structure and Identity

Recombinant Human CD5L (also known in literature as CD5 antigen-like; AIM/Api6) is a secreted, cysteine-rich glycoprotein produced recombinantly and supplied with a polyhistidine tag.

• SKU: rp144064
• Product name: Recombinant Human CD5L Protein
• Category: Protein (recombinant; human sequence)
• Tag: His tag (position not specified)
• Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
  • Typical literature range: ~35–45 kDa for mature, glycosylated monomer depending on glycoform (literature)
• SMILES/InChI/InChIKey: Not applicable to proteins; not specified for this item.
• Structural features (literature):
  • Member of the scavenger receptor cysteine-rich (SRCR) superfamily; contains multiple SRCR domains rich in disulfide bonds.
  • Secreted, N-glycosylated protein; disulfide-stabilized tertiary structure.
• 2D/sequence-level description (literature):
  • Linear polypeptide chain corresponding to human CD5L ectodomain; the His-tag facilitates IMAC purification and detection. Exact sequence length, tag orientation, and linker details are not specified for this item.

Synthetic Utility

Not applicable in the classical synthetic chemistry sense. Recombinant Human CD5L Protein is a biologic macromolecule rather than a small-molecule building block or catalyst.

Relevance to laboratory workflows (general):

• As a capture/bait protein in chemical proteomics to identify interacting small molecules or proteins under native-like conditions.
• As a standard in bioanalytical method development (e.g., LC-MS quantification of CD5L in complex matrices post-immunoenrichment).
• As a functional component in biomaterials or surface-functionalized platforms (e.g., NTA-coated biosensors) for binding studies.

For synthetic organic transformations, see other catalog sections; for this protein, focus on the Application Protocols and Reaction Conditions tabs tailored to biochemical assays.

Target Specificity

Not applicable. This product is a recombinant human protein (CD5L) and not an antibody or affinity reagent. No antigen/epitope specificity, clone information, isotype, or species reactivity data apply.