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N-terminal Sequence Analysis
Thr39
State of Matter
Lyophilized
Predicted Molecular Mass
The predicted molecular weight of Recombinant Human CXCL2, GROβ, MIP-2, CINC3, is Mr 7.5 kDa.
Predicted Molecular Mass
7.5
Formulation
This recombinant protein was lyophilized from a 0.2 μm filtered solution in 35% acetonitrile (CH3CN) and 0.1% trifluoroacetic acid (TFA).
Storage and Stability
This lyophilized protein is stable for six to twelve months when stored desiccated at -20°C to -70°C. After aseptic reconstitution, this protein may be stored at 2°C to 8°C for one month or at -20°C to -70°C in a manual defrost freezer. Avoid Repeated Freeze Thaw Cycles. See Product Insert for exact lot specific storage instructions.
Applications and Recommended Usage ? (Quality Tested by Leinco)
ELISA Sandwich: This antibody is useful as the capture antibody in a sandwich ELISA. The suggested coating concentration is 5 µg/ml (100 µl/well) µg/ml. Flow Cytometry: PN:A106 Flow Cytometry: It is recommended to use the indirect method for signal enhancement when enumerating cells expressing CXCR5. A suggested method would be to stain cells expressing CXCR5 with approximately 10 µl per test. A typical test sample constitutes approximately 50 µl of packed whole blood or 1 x 105 continuous passage or activated cell cultures that have been centrifuged at 500 X g for five minutes. Labeling of the cells with the biotin conjugate should be followed by PN:A104, resuspended in 200-400 µl of 1X PBS.
Leinco Protein Advisor
Powered by AI: AI is experimental and still learning how to provide the best assistance. It may occasionally generate incorrect or incomplete responses. Please do not rely solely on its recommendations when making purchasing decisions or designing experiments.
Recombinant Human GROβ/CXCL2 (aa 39-107) is used in research applications primarily for its potent ability to recruit and activate neutrophils and other leukocytes, making it a valuable tool for studying inflammation, immune cell migration, hematopoietic stem cell mobilization, and tumor microenvironment dynamics.
Key scientific applications and rationale include:
Neutrophil and Basophil Recruitment: GROβ/CXCL2 is a strong chemoattractant for neutrophils and basophils, mediating their migration to sites of tissue injury or infection. This property is essential for modeling acute inflammatory responses and studying leukocyte trafficking in vitro and in vivo.
Hematopoietic Stem Cell Mobilization: GROβ/CXCL2 can rapidly mobilize hematopoietic stem and progenitor cells (HSPCs) into the peripheral blood, with evidence showing it mobilizes more primitive and competitive HSCs compared to G-CSF, leading to accelerated hematopoietic recovery in transplantation models. This makes it useful for research on stem cell biology and transplantation.
Tumor Microenvironment Studies: CXCL2 is a key player in the tumor microenvironment, influencing tumor progression by promoting immune cell infiltration, particularly neutrophils, and modulating tumor cell migration and invasion. It is used to dissect mechanisms of tumor-immune interactions and metastasis.
CXCR2 Ligand Studies: GROβ/CXCL2 binds with high affinity to the chemokine receptor CXCR2, making it suitable for receptor binding assays, signaling studies, and screening of CXCR2-targeted therapeutics.
Inflammation and Immunology Research: As an ELR+ CXC chemokine, GROβ/CXCL2 is inducible by inflammatory mediators (e.g., IL-1, TNFα) and is involved in the regulation of immune responses, making it a model chemokine for studying cytokine networks and inflammatory signaling pathways.
Bioactivity and Functional Assays: The aa 39-107 fragment corresponds to the mature, biologically active form of GROβ, ensuring robust and reproducible results in chemotaxis, cell activation, and myeloperoxidase release assays.
Summary of best practices:
Use the mature aa 39-107 form for functional studies, as it reflects the active chemokine produced in vivo.
GROβ/CXCL2 is suitable for in vitro and in vivo assays involving leukocyte migration, stem cell mobilization, and tumor biology.
It is a critical reagent for dissecting CXCR2-mediated signaling and immune cell dynamics in health and disease.
In summary, Recombinant Human GROβ/CXCL2 (aa 39-107) is a versatile and well-characterized reagent for research in immunology, stem cell biology, and cancer, owing to its potent chemotactic and cell-activating properties.
Recombinant Human GROβ/CXCL2 (aa 39-107) can be used as a standard for quantification or calibration in ELISA assays, provided it is properly validated for this purpose and matches the epitope recognized by your assay antibodies.
Key considerations and supporting details:
ELISA standards are typically purified recombinant proteins that match the analyte being measured. The aa 39-107 fragment corresponds to the mature form of GROβ/CXCL2, which is commonly used in commercial ELISA kits for calibration.
Many commercial ELISA kits for GROβ/CXCL2 use recombinant protein standards covering aa 39-107, indicating suitability for quantification. These standards are used to generate a standard curve, which is essential for accurate quantification of GROβ/CXCL2 in samples.
It is important to ensure that the recombinant protein is of high purity, properly reconstituted, and that its concentration is accurately determined before use. Carrier proteins such as BSA may be added to enhance stability, but carrier-free versions are available if BSA interferes with your assay.
The recombinant protein should be validated for use as a standard in your specific ELISA system. This includes confirming that the antibodies in your assay recognize the same epitope as the recombinant standard and that the standard curve generated is linear and reproducible.
Some recombinant proteins are intended for bioassays rather than ELISA calibration; always check the product specifications to confirm intended use. For ELISA, select a recombinant protein that is specifically recommended or validated as a standard.
Best practices:
Prepare serial dilutions of the recombinant GROβ/CXCL2 (aa 39-107) to generate a standard curve for your ELISA.
Validate the standard curve for accuracy, precision, and linearity in your assay system.
Store and handle the recombinant protein according to manufacturer recommendations to maintain stability and activity.
In summary, recombinant Human GROβ/CXCL2 (aa 39-107) is suitable as an ELISA standard if it is properly validated and matches the assay’s antibody specificity. Always confirm compatibility with your specific assay protocol.
Recombinant Human GROβ/CXCL2 (aa 39-107) has been validated primarily for bioactivity assays, especially those measuring chemotactic and neutrophil-activating functions, in published research.
Key validated applications include:
Bioassays for chemotaxis: The protein is widely used to induce chemotaxis (directed cell migration) in various cell types, particularly neutrophils and cells expressing the CXCR2 receptor. For example, it has been shown to induce chemotaxis of BaF3 mouse pro-B cells transfected with human CXCR2, with a reported ED₅₀ of 1–5 ng/mL.
Neutrophil activation assays: It is validated for inducing myeloperoxidase release from cytochalasin B-treated human neutrophils, a standard measure of neutrophil activation. The ED₅₀ for this effect is 0.1–0.3 µg/mL.
General bioactivity in cell culture: The protein is used in cell culture systems to study leukocyte recruitment, activation, and signaling, as well as in assays involving basophil activation.
Binding assays: It has been used in binding assays to characterize receptor interactions, such as with CXCR2.
Functional studies in immunology and cancer research: Published studies have used recombinant CXCL2 to investigate neutrophil recruitment, macrophage-mediated immune responses, and tumor microenvironment modulation.
Summary Table of Validated Applications
Application Type
Description/Assay Example
Reference
Chemotaxis/Bioassay
Migration of CXCR2+ cells, neutrophil chemotaxis
Neutrophil Activation
Myeloperoxidase release from human neutrophils
Cell Culture Bioactivity
Leukocyte/basophil recruitment and activation
Binding Assay
Receptor (CXCR2) binding studies
Immunology/Cancer Research
Modulation of immune cell recruitment in disease models
Additional Notes:
The protein is typically produced in E. coli and corresponds to the mature form (Thr39-Asn107) of human CXCL2.
It is not validated for diagnostic or therapeutic use, but is a standard reagent for research on chemokine signaling, inflammation, and tumor immunology.
Published research has used this protein to study neutrophil recruitment in infection and cancer, as well as to dissect chemokine-receptor interactions in vitro.
If you require details on a specific assay protocol or a particular disease model, please specify for more targeted information.
To properly reconstitute and prepare Recombinant Human GROβ/CXCL2 (aa 39–107) for cell culture experiments, follow these best practices based on manufacturer recommendations and scientific protocols:
Reconstitution Steps
Centrifuge the Vial Briefly centrifuge the lyophilized protein vial in a microcentrifuge (20–30 seconds) before opening to ensure all powder is at the bottom.
Reconstitution Buffer Reconstitute the lyophilized protein in sterile PBS (phosphate-buffered saline) or sterile distilled water.
Typical concentration: 100 µg/mL (as recommended by most suppliers).
For carrier-free proteins, use sterile PBS or water.
If using a carrier protein (e.g., for stability), add 0.1–1% BSA (bovine serum albumin) or HSA (human serum albumin) to the buffer.
Reconstitution Procedure
Gently pipette the buffer down the side of the vial to avoid foaming.
Allow several minutes for complete dissolution (do not vortex).
Mix gently by swirling or inverting the vial.
Aliquoting
Prepare working aliquots to avoid repeated freeze-thaw cycles.
Store aliquots at –20°C or –80°C for long-term storage.
Storage and Stability
Lyophilized protein: Store at 2–8°C (desiccated) until reconstitution.
Reconstituted protein:
For short-term use: 2–8°C for up to 1 week (with carrier protein) or 1 month (with carrier protein, per some suppliers).
For long-term storage: ≤ –20°C (preferably –80°C) for up to 6 months (with carrier protein).
Avoid repeated freeze-thaw cycles.
Preparation for Cell Culture
Dilute the reconstituted stock to the desired working concentration in cell culture medium or assay buffer.
Typical working concentrations for chemotaxis or bioassays: 10–100 ng/mL.
For optimal activity, ensure the protein is fully dissolved and aliquoted to minimize degradation.
Additional Tips
Confirm protein concentration and integrity by running a small sample on SDS-PAGE if needed.
Use a manual defrost freezer for storage to prevent temperature fluctuations.
Summary Protocol
Step
Action
1
Centrifuge vial before opening
2
Reconstitute in sterile PBS or water to 100 µg/mL (add 0.1–1% BSA if desired)
3
Gently mix, do not vortex
4
Aliquot and store at ≤ –20°C (or –80°C)
5
For cell culture, dilute to working concentration (e.g., 10–100 ng/mL) in medium
This protocol ensures optimal activity and stability of Recombinant Human GROβ/CXCL2 (aa 39–107) for cell culture experiments.
Products are for research use only. Not for use in diagnostic or therapeutic procedures.
