Detection Method
Sandwich ELISA
Intended Use
Mouse M-CSF ELISA development kit contains the key components required for the quantitative measurement of natural and/or recombinant M-CSF in a sandwich ELISA format within the range of 32 -1,000 pg/mL. Using the ELISA protocol described below, this kit provides sufficient reagents to assay M-CSF in approximately 1,500 ELISA plate wells.
Contents of Kit
1. PBS: Dilute 10X PBS to 1X PBS, pH 7.2 in sterile water.
2. Wash Buffer: 0.05% Tween-20 in PBS
3. Block Buffer: 1.0% BSA in PBS
4. Diluent: 1.0% BSA in PBS
5. ELISA microplates (Thermo Fisher Cat. # 456529);
6. BSA (Sigma Cat. # A-7030);
7. Stop Solution 2 M Sulfuric Acid (Sigma Cat. # 339741);
8. Dulbecco's PBS [10X] (Gibco BRL Cat. # 14200-075).
Storage
Store the kit at 4°C upon receipt. For more detailed information, please download the following document on our website.
General Description
Macrophage colony-stimulating factor, or M-CSF, is a secreted cytokine which influences hemopoietic stem cells to differentiate into macrophages or other related cell types. M-CSF is a key regulator of cellular proliferation, differentiation, and survival of blood monocytes, tissue macrophages and their progenitor cells. It enhances cytotoxicity, superoxide production, phagocytosis, chemotaxis, and secondary cytokine production in monocytes and macrophages. It binds to the Colony stimulating factor 1 receptor.
Reconstitution And Storage
##Capture Antibody: ## 360μg of antigen-affinity purified goat anti-M-CSF. Centrifuge vial prior to opening. Reconstitute in 1.0 ml PBS for a concentration of 360μg/ml. Following reconstitution the Capture antibodies may be stored at 2-8°Cfor up to 6 months. For long term storage it is recommended to aliquot into working volumes and store at -70°C in a manual defrost freezer.
##Detection Antibody: ##18 μg of biotinylated antigen-affinity purified goat anti-M-CSF. Centrifuge vial prior to opening. Reconstitute in 1.0 ml sterile 1.0% BSA in PBS for a concentration of 18 μg/ml. Following reconstitution the Detection antibodies may be stored at 2-8°C for up to 6 months. For long term storage it is recommended to aliquot into working volumes and store at -70°C in a manual defrost freezer.
##Mouse M-CSF Standard: ##25 ng of recombinant M-CSF. Centrifuge vial prior to opening. Reconstitute in 0.5 ml sterile 1.0% BSA in PBS for a concentration of 50 ng/ml. The Standard may be stored at 2-8°C for one month or aliquoted and stored at -70°C for up to three months in a manual defrost freezer.
##UltraAvidin-HRP Conjugate: ##40μl vial. Upon receipt, UltraAvidin-HRP conjugate should be stored at 2-8°C, DO NOT FREEZE.
##TMB Liquid Substrate: ##Aspirate and wash plate 4 times. Add 100μl of TMB HRPO Microwell Substrate Standard Kinetic One Component "Ready Use" to each well. Incubate at room temperature and monitorcolor development with an ELISA plate reader at 450 nm with wavelength correction set at 540 nm or 570 nm. The reaction may be stopped after 15-20 minutes by adding 100μl of 2 M sulfuric acid to each well.
Citations
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