Tough cryogels were fabricated using a combination of covalent and ionic crosslinking of alginate. First, covalently crosslinked-only cryogels were formed.[14 (link)] Briefly, 1.5% (w/v) MA-alginate solution in deionized water was mixed with polymerization initiators TEMED (0.5% w/v) and APS (0.25% w/v). The mixture was quickly added to Teflon molds (5 mm diameter, 1 mm height) which were placed at −20°C overnight while crosslinking occurred. Cryogels were thawed and then soaked in 200 mM calcium chloride to form ionic crosslinking with calcium ions. They were soaked for different durations to vary calcium concentration in the gels. To quantify calcium concentrations in the gels, the gels were washed in a 25 mM HEPES buffer supplemented by 2 mM CaCl2 and 140 mM NaCl for 1 min under gentle stirring to remove excess calcium ions from the pores. Gels were then digested in 1 mL 10 U/mL alginate lyase on a shaker by incubating overnight at RT. Samples were quantified using a calcium assay kit (BioVision, Milpitas, CA). When used, GM-CSF, CpG-ODN and OVA, were mixed in the MA-alginate polymer solution prior to cryogelation. Cell-adhesive tough cryogels were fabricated by adding 0.8% (w/v) of ACRL-PEG-RGD to the MA-alginate solution as a comonomer prior to cryogelation.
Calcium assay kit
Manufactured by Abcam
Sourced in United Kingdom, United States
The Calcium Assay Kit is a colorimetric assay for the quantitative determination of calcium levels in various sample types, including biological fluids, cell lysates, and tissue homogenates. The kit utilizes a chromogenic complex that changes color in the presence of calcium, allowing for the accurate measurement of calcium concentrations in the sample.
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Lab products found in correlation
Phosphate assay kit, by Abcam (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
A25oh vitamin d elisa assay kit, by Eagle Biosciences (2 mentions)
Fluo 4 am, by Thermo Fisher Scientific (2 mentions)
Zinc assay kit, by Abcam (1 mentions)
Bovine fetuin a, by Merck Group (1 mentions)
Synergy 2, by Agilent Technologies (1 mentions)
Infinite m200 pro, by Tecan (1 mentions)
Glucose assay kit, by Abcam (1 mentions)
Human albumin elisa kit, by Abcam (1 mentions)
Human il 6 elisa kit, by Abcam (1 mentions)
Human il 17 elisa kit, by Abcam (1 mentions)
Amylase assay kit, by Abcam (1 mentions)
Human c reactive protein elisa kit, by Abcam (1 mentions)
Human tnf alpha elisa kit, by Abcam (1 mentions)
Human il 10 elisa kit, by Abcam (1 mentions)
Xt 2000i, by Sysmex (1 mentions)
Alkaline phosphatase assay kit, by Abcam (1 mentions)
Puromycin, by Thermo Fisher Scientific (1 mentions)
Hippocalcin, by Abcam (1 mentions)
23 protocols using calcium assay kit
1
Tough Cryogels via Covalent and Ionic Crosslinking
2
Serum Biomarker Quantification in Mice
3
Colorimetric Ca2+ Quantification in Spleen Cells
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The serum was subjected to a colorimetric method for the detection of the Ca2+ concentration according to the manufacturer’s protocol (Calcium Assay Kit, BioVision).
To detect the Ca2+ concentration in spleen cells, the fresh spleen tissue was made into a single cell suspension (2 x 106 cells per EP tube). The cells were resuspended in serum-free medium at a final concentration of 2.5 μmol/L of Fluo 3-Am. The suspension was placed at 37°C at 5% CO2 for 30 min. The suspension was centrifuged at 1500 rpm for 5min, and the supernatant was discarded. The cells were washed with calcium-free PBS twice and suspended in 500 μl calcium-free PBS. The Ca2+ concentration of the spleen cells was detected by flow cytometry.
To detect the Ca2+ concentration in spleen cells, the fresh spleen tissue was made into a single cell suspension (2 x 106 cells per EP tube). The cells were resuspended in serum-free medium at a final concentration of 2.5 μmol/L of Fluo 3-Am. The suspension was placed at 37°C at 5% CO2 for 30 min. The suspension was centrifuged at 1500 rpm for 5min, and the supernatant was discarded. The cells were washed with calcium-free PBS twice and suspended in 500 μl calcium-free PBS. The Ca2+ concentration of the spleen cells was detected by flow cytometry.
4
Serum Biomarker Quantification in Mice
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Blood samples were collected from WT and Osx-Nf1 KO mice at sacrifice. Vitamin
D, phosphate and calcium concentration in mouse serum was determined using a
25OH-Vitamin-D ELISA Assay kit (Eagle Biosciences, cat# VID31-K01), a Phosphate
Assay kit (BioVision, cat # k410–500) and a Calcium Assay kit (BioVision,
cat# k380–250), respectively, according to the manufacturer’s
instructions.
D, phosphate and calcium concentration in mouse serum was determined using a
25OH-Vitamin-D ELISA Assay kit (Eagle Biosciences, cat# VID31-K01), a Phosphate
Assay kit (BioVision, cat # k410–500) and a Calcium Assay kit (BioVision,
cat# k380–250), respectively, according to the manufacturer’s
instructions.
5
Quantifying Cellular and Tissue Ions
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Following kits were used for determination of concentrations of calcium, iron, and zinc ions: Calcium Assay Kit (Colorimetric) (Abcam, ab102505), Iron Assay Kit (Colorimetric) (Abcam, ab83366), and Zinc Assay Kit (Abcam, ab102507), respectively. In cellular analyses, 6 × 105 cells were passaged in 10 cm diameter-plates and allowed to attach overnight. Cells were then harvested and transferred to tubes, washed with the PBS buffer (137 mM NaCl; 2.7 mM KCl; 10 mM Na2HPO4; 1.8 mM KH2PO4), and centrifuged (300 x g for 10 min). The experiment was performed in 3 independent replications. In animals tissues analysis, 10 mg of brain, liver or spleen were separated and washed with the PBS buffer. Cellular pellets and tissues were next lysed/homogenized for 3 h with Calcium Assay Buffer, Iron Assay Buffer or EDTA-free lysis buffer (0.9% NaCl; 0.5% Triton X-100; 0.1% SDS; 1% sodium deoxycholate; 50 mM Tris-HCl pH 7.5) to determine concentrations of calcium, iron or zinc ions, respectively. To obtain cellular lysate or tissue homogenate, samples were centrifuged at 16,000 x g for 10 min. Calcium, iron or zinc ion concentrations were assessed by colorimetric measurements and appropriate calculations, according to the manufacturer’s instructions.
6
Serum Biomarker Quantification Protocol
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Blood samples were collected from the lateral tail veins and allowed to clot at room temperature protected from light, followed by centrifugation at 1,500g for 10 min at 4°C. Supernatant serums were collected. 25(OH)D level was detected using 25(OH)D ELISA kit (Immunodiagnostic Systems); calcium and phosphate levels were detected using calcium assay kit and phosphate assay kit (both Abcam) according to the manufacturer's instructions.
7
Intracellular Calcium Quantification
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To measure intracellular Ca2+ levels, the Calcium Assay Kit from Abcam was used according to the manufacturer’s protocols.
8
Intracellular Calcium Quantification in Neuropathic Pain
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Calcium Assay Kit (ab102505, Abcam) was used to quantify the intracellular calcium content in hind paw and lumbar spinal cord tissues, according to the manufacturer’s instructions.
9
Synthesis and Characterization of Calcium Phosphate Particles
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CPP were synthesized as previously reported6 (link),21 (link). Briefly, a CPP solution containing 1 mg/mL bovine fetuin-A (Sigma-Aldrich, St. Louis, MO, USA), 10 mM CaCl2, 6 mM Na2HPO4, 140 mM NaCl, and 50 mM Tris–HCL (pH 7.4) was gently shaken and incubated at 37 °C for at least 24 h. Synthetic CPP were separated using a spin-filtered membrane cartridge with a 300 kDa cut-off (Sartorius AG, Göttingen, Germany). CPP concentrations were determined by bicinchoninic acid (BCA) assay as a protein level. The ionic calcium content of secondary CPP was also measured by Calcium Assay Kit (Cat No. ab102505, abcam, Cambridge, UK). One hundred μg/mL of secondary CPP as a protein concentration included 45 μg/mL of ionic calcium (Supplementary Fig. 8 ). Synthetic secondary CPP were used for experiments from 24 to 168 h after the first mixture of the solution.
10
Intracellular Ca2+ Measurement in Osteoclasts
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Intracellular Ca2+ was measured using a Calcium Assay Kit (#ab102505, Abcam, Cambridge, MA, USA) according to the manufacturer’s protocol. In brief, after 6-days of osteoclast differentiation with M-CSF and sRANKL in the absence or presence of TA, GA, or EA (30 μM), cells were harvested and homogenized with calcium assay buffer. 50 µL of cell lysate was added to each well of a 96-wells plate. 90 µL of the Chromogenic Reagent was added to each well, followed by 60 µL of Calcium Assay Buffer. Samples were mixed and incubated at RT for 10 min in the dark. A standard curve was prepared using the standard dilutions. Absorbance was recorded at 575 nm using a microplate reader (Synergy 2, BioTek Instruments, Inc., Winooski, VT, USA). Each experiment was performed in triplicate, and experiments were performed at least three times.
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