Immunization grade bovine type II collagen (Chondrex Inc., Redmond, USA) was dissolved in 0.1M glacial acetic acid (Jindongtianzheng Precision Chemical Reagent Factory, Tianjin, China) at the concentration of 2 mg/ml and stirred overnight at 4°C. It was then mixed with an equal amount of Freund's complete adjuvant (Sigma-Aldrich, St. Louis, USA) and emulsified with a homogenizer on ice. When the emulsion drop floats completely on the surface of the water without spreading out, it indicates complete emulsification [15 (link), 16 (link)].
Immunization grade bovine type 2 collagen
Manufactured by Chondrex
Sourced in United States
Immunization grade bovine type II collagen is a laboratory reagent used for research purposes. It is derived from bovine cartilage and is of high purity and quality, suitable for use in immunological studies and experiments.
Automatically generated - may contain errors
Lab products found in correlation
Complete freund s adjuvant, by Merck Group (1 mentions)
Incomplete freund s adjuvant, by Chondrex (1 mentions)
Ab22510, by Abcam (1 mentions)
Ab8226, by Abcam (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Gotaq qpcr master mix, by Promega (1 mentions)
Complete freund s adjuvant, by Chondrex (1 mentions)
Cpg odn 1826 tlrl 1826, by InvivoGen (1 mentions)
Easysep human pan dc pre enrichment kit, by STEMCELL (1 mentions)
Bovine type 2 collagen, by Chondrex (1 mentions)
6 protocols using immunization grade bovine type 2 collagen
1
Emulsification of Bovine Collagen in Freund's Adjuvant
2
Collagen-Induced Arthritis Treatment Evaluation
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Immunization grade bovine type II collagen (Chondrex, USA) was emulsified with an equal volume of incomplete Freund’s adjuvant. On day 0, 0.2 mL of the emulsion was subcutaneously injected into the tail base of the treatment group rats, followed by a booster injection on day 7. Treatment group rats were given oral gavage once a day. After administration, six rats in each group were selected for each efficacy index on days 15th and 28th.
Sodium carboxymethyl cellulose (CMC-Na) was used as a vehicle for drug and control treatments. After the onset of arthritis, the rats were divided into the following six groups (n = 12); normal (CMC-Na); vehicle-treated (CIA with CMC-Na); methotrexate (MTX)-treated [(CIA with 0.2 mg/kg by weight (bw)]; low concentration (CIA with 1 mg/kg bw) LY411575-treated group; medium concentration (CIA with 5 mg/kg bw) LY411575-treated group; high concentration (CIA with 10 mg/kg bw) LY411575-treated group. Rats were administered the treatments by oral gavage once a day, and six rats from each group were tested for treatment efficacy on days 15 and 28, respectively.
Sodium carboxymethyl cellulose (CMC-Na) was used as a vehicle for drug and control treatments. After the onset of arthritis, the rats were divided into the following six groups (n = 12); normal (CMC-Na); vehicle-treated (CIA with CMC-Na); methotrexate (MTX)-treated [(CIA with 0.2 mg/kg by weight (bw)]; low concentration (CIA with 1 mg/kg bw) LY411575-treated group; medium concentration (CIA with 5 mg/kg bw) LY411575-treated group; high concentration (CIA with 10 mg/kg bw) LY411575-treated group. Rats were administered the treatments by oral gavage once a day, and six rats from each group were tested for treatment efficacy on days 15 and 28, respectively.
3
Quantitative Analysis of Doxorubicin Hydrochloride
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PEGylated doxorubicin hydrochloride liposome injections were obtained from Changzhou Kinyond Manufacturing Co., Ltd. (Jintan, Jiangsu, China). Doxorubicin hydrochloride for injection was purchased from Zhejiang Hisun Pharmaceutical Co., Ltd. (Taizhou, Zhejiang, China). Immunization-grade bovine type II collagen, complete Freund’s adjuvant containing 2 mg/mL M. tuberculosis, and incomplete Freund’s adjuvant were purchased from Chondrex, Inc (Redmond, WA, USA). Other chemicals were of analytical grade and obtained from commercial sources.
Doxorubicin hydrochloride and daunorubicin hydrochloride (internal standard, IS) were purchased from the National Institutes for Food and Drug Control (Beijing, China). Acetonitrile and methanol were of HPLC grade and purchased from Tianjin Shield Fine Chemicals Co., Ltd (Tianjin, China). Ammonium acetate and glacial acetic acid were of analytical grade and obtained from Tianjin Hengxing Chemical Reagent Co., Ltd (Tianjin, China). Ultrapure water was obtained using a Millipore Milli-Q Academic system (Millipore Corporation, Bedford, MA, USA).
Doxorubicin hydrochloride and daunorubicin hydrochloride (internal standard, IS) were purchased from the National Institutes for Food and Drug Control (Beijing, China). Acetonitrile and methanol were of HPLC grade and purchased from Tianjin Shield Fine Chemicals Co., Ltd (Tianjin, China). Ammonium acetate and glacial acetic acid were of analytical grade and obtained from Tianjin Hengxing Chemical Reagent Co., Ltd (Tianjin, China). Ultrapure water was obtained using a Millipore Milli-Q Academic system (Millipore Corporation, Bedford, MA, USA).
4
Osthole Attenuates Rheumatoid Arthritis
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OMT was purchased from Ningxia Bauhinia Pharmacy Co., Ltd. (Ningxia, China). OMT 100, 50 and 25 mg/kg (dissolved in normal saline) was administrated via intraperitoneal injection (i.p.). Immunization grade bovine type II collagen and complete Freund's adjuvant were purchased from Chondrex, Inc. (Redmond, WA, USA). Enzyme-linked immunosorbent assay (ELISA) kits for IL-17 and TNF-α, and mouse monoclonal antibodies against FOXP3 (ab22510), RORγt (ab41942) and β-actin (ab8226) were purchased from Abcam (Cambridge, MA, USA). TRIzol was obtained from Gibco; Thermo Fisher Scientific, Inc. (Waltham, MA, USA). A reverse transcription kit was purchased from TransGen Biotech, Inc. (Beijing, China). Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis was performed using GoTaq® qPCR master mix (Promega Corporation, Madison, WI, USA). BCA and enhanced chemiluminescence (ECL) kits were from Pierce, Thermo Fishers Scientific, Inc.
5
Purification and Immunization Protocol
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All flow antibodies were purchased from BD Biosciences and BioLegend of USA. And all ELISA kits were purchased from NEO Biosciences of China. EasySep™ Human Pan-DC Pre-Enrichment Kit for DC purification was purchased from STEMCELL of Canada and the purity of the DC up to 86.6 % in the research (Fig. S1). Immunization Grade Bovine Type II Collagen, immunization Grade Chick Type II Collage, Complete and incomplete Freund's adjuvants were purchased from Chondrex, USA. CpG ODN 1826 (tlrl-1826) and recombinant murine GM-CSF (AF-315-03) were purchased from InvivoGen and PeproTech of USA.
6
Mouse Model of Collagen-Induced Arthritis
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After 1 week of housing, mice were sensitized by subcutaneous injection in their tails of 100 µg bovine type II collagen in an adjuvant solution for primary sensitization. The collagen adjuvant solution was prepared by the following method.
Immunization grade bovine type II collagen (Chondrex) was dissolved at a concentration of 4 mg/mL in buffer (20 mM Tris-HCl, pH 8.0 and 150 mM NaCl) and stirred under dark conditions at 4°C overnight. The bovine type II collagen solution was emulsified with an equal amount of complete Freund's adjuvant (CFA, 4 mg/ml, Chondrex), which was mixed on ice in cold room (4°C) for 15 min. Two weeks after primary sensitization, mice were secondary sensitized using the same injection procedure with 200 µg bovine type II collagen in adjuvant solution. bovine type II collagen was dissolved at a concentration of 8 mg/mL in buffer (20 mM Tris-HCl, pH 8.0 and 150 mM NaCl) and stirred in the dark at 4°C overnight. The bovine type II collagen solution was emulsified with an equal amount of incomplete Freund's adjuvant (IFA, Chondrex), which was mixed on ice for 10 min.
Immunization grade bovine type II collagen (Chondrex) was dissolved at a concentration of 4 mg/mL in buffer (20 mM Tris-HCl, pH 8.0 and 150 mM NaCl) and stirred under dark conditions at 4°C overnight. The bovine type II collagen solution was emulsified with an equal amount of complete Freund's adjuvant (CFA, 4 mg/ml, Chondrex), which was mixed on ice in cold room (4°C) for 15 min. Two weeks after primary sensitization, mice were secondary sensitized using the same injection procedure with 200 µg bovine type II collagen in adjuvant solution. bovine type II collagen was dissolved at a concentration of 8 mg/mL in buffer (20 mM Tris-HCl, pH 8.0 and 150 mM NaCl) and stirred in the dark at 4°C overnight. The bovine type II collagen solution was emulsified with an equal amount of incomplete Freund's adjuvant (IFA, Chondrex), which was mixed on ice for 10 min.
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