Twenty-four male DBA/1 mice, age of four weeks, weighing ~20 g, were purchased from Beijing Vital River Laboratory Animal Technology Limited. Immunization grade bovine type II collagen (lyophilized), Freund's incomplete adjuvant, and Freund's complete adjuvant were purchased from Chondrex (WA, USA). ELISA kit was purchased from BlueGene (Shanghai, China). NLRP3 monoclonal antibodies were purchased from Abcam (Cambridge, UK). Anti-mouse two-step immunohistochemical detection kit was purchased from Zhongshan Golden Bridge Biotechnology Company (Beijing, China).
Incomplete freund s adjuvant
Manufactured by Chondrex
Sourced in United States, China
Incomplete Freund's adjuvant is a laboratory reagent used to enhance the immune response in experimental settings. It consists of a water-in-oil emulsion without the addition of killed or attenuated microorganisms. The core function of Incomplete Freund's adjuvant is to stimulate the immune system, which can be useful in various immunological studies and assays.
Automatically generated - may contain errors
Lab products found in correlation
Complete freund s adjuvant, by Chondrex (44 mentions)
Bovine type 2 collagen, by Chondrex (34 mentions)
Type 2 collagen, by Chondrex (6 mentions)
Bovine type 2 collagen cii, by Chondrex (6 mentions)
Chicken cii, by Chondrex (5 mentions)
Chicken type 2 collagen, by Chondrex (4 mentions)
Dba 1 mice, by Orient Bio (4 mentions)
Trizol, by Thermo Fisher Scientific (3 mentions)
Complete freund s adjuvant cfa, by Chondrex (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Complete freund s adjuvant, by Merck Group (2 mentions)
Mycobacterium tuberculosis h37ra, by Chondrex (2 mentions)
Chick type 2 collagen, by Chondrex (2 mentions)
Hplc grade methanol, by Merck Group (2 mentions)
Dba 1j mice, by Central Lab Animal (2 mentions)
Bovine type 2 collagen solution, by Chondrex (2 mentions)
Bovine cii, by Chondrex (2 mentions)
Dba 1j mice, by Orient Bio (2 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (2 mentions)
Dba 1j mice, by Shanghai Laboratory Animal Center (1 mentions)
83 protocols using incomplete freund s adjuvant
1
Murine Model of Collagen-Induced Arthritis
2
Collagen-Induced Arthritis Model with MSC Therapy
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CIA was induced in male DBA/1J mice (age 7–9 weeks) purchased from Shanghai Laboratory Animal Center (SLAC, Shanghai, China). The mice (n = 20) were maintained under standard conditions in our university's central animal laboratory and randomly placed in cages. Bovine type II collagen (CII; Chondrex) was dissolved in 50 mM acetic acid and emulsified with an equal volume of Freund's complete adjuvant (Sigma-Aldrich). The mice were immunized at the base of the tail with 100 μL emulsion containing 200 μg CII. After 21 days, the mice were administered a booster dose of 100 μg CII (2 mg/mL) emulsified with Freund's incomplete adjuvant (Chondrex) via intradermal injection into the tail. On days 28, 32, and 38 after the first immunization, the mice were anesthetized and 106 MSCs in 7 μL PBS were injected intra-articularly into the right knee for the SM-MSC treatment group (n = 8); the control mice received 7 μL PBS intra-articularly (n = 8).
3
Collagen-Induced Arthritis in Mice
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CIA was induced according to the previous methods [27 (link)]. Mice were immunized intradermally at the base of tail with 100 μg of type II collagen (Chondrex, WA, USA) emulsified with an equal volume of Freund's complete adjuvant (Chondrex, WA, USA) and intraperitoneally boosted with type II collagen (100 μg in 0.05 M acetic acid) emulsified with an equal volume of Freund's incomplete adjuvant (Chondrex, WA, USA) at 23 days after the initial immunization. In intact control mice, only vehicle, 0.05 M acetic acid was intradermally injected instead of collagen immunization and intraperitoneally administered instead of antigen boosting, respectively.
4
Modeling Autoimmune Arthritis in Mice
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Following one week of acclimatization feeding, SPF DBA/1 mice (male, six-week-old, 18-20 g) were randomly divided into two groups (n = 6) as follows: the model group (MG, type II bovine collagen + Freund’s complete adjuvant/Freund’s incomplete adjuvant) and NC group (CG, saline). On day 0, the mice in the MG were subcutaneously injected with 100 μL of 1:1 type II bovine collagen and Freund’s complete adjuvant (Chondrex, Redmond, WA, USA) at multiple points of the caudal root. After 21 days, the same mice were injected with 1:1 type II bovine collagen + Freund’s incomplete adjuvant (Chondrex, Redmond, WA, USA). As per established scoring criteria [58 (link)], a score > 4 was considered to indicate successful modeling. Following successful modeling, peripheral blood was obtained from the mice, and PBMCs were isolated according to the instructions of the whole blood single nucleated cell isolation solution (Solarbio, Beijing, China). Total RNA was extracted by using TRIzol (Thermo Fisher, Waltham, MA, USA). This study was approved by the Guangzhou Medical University Ethics Committee.
5
Collagen-Induced Arthritis in Mice
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Male C57BL/6 wild‐type mice were purchased from the Laboratory Animal Center of Southern Medical University. C57BL/6 Gsdme‐knockout (Gsdme−/−) mice were generated by CRISPR/Cas‐mediated genome engineering (Cyagen Bioscience). CIA was induced in 8‐week‐old male wild‐type and Gsdme−/− mice. Chicken type II collagen (catalog no. 20012; Chondrex) was dissolved in 0.05M acetic acid (final concentration 2 mg/ml) and then emulsified with Freund's complete adjuvant (CFA) containing heat‐killed Mycobacterium tuberculosis H37RA (Chondrex). C57BL/6 mice were injected intradermally with a 100‐μl emulsion at the base of tail on days 1 and 21 (33 (link)). Clinical assessment of CIA was performed daily for each limb as previously described (34 (link)).
CIA was generated in DBA/1J mice as previously described (35 (link)). Eight‐week‐old male DBA/1J mice were immunized with 100 mg bovine type II collagen (2 mg/ml; Chondrex) emulsified 1:1 in CFA (containing 1 mg/ml M tuberculosis; Chondrex) on day 1 and then injected with Freund's incomplete adjuvant (Chondrex) on day 21.
All animals were maintained under specific pathogen–free conditions in the Laboratory Animal Center of Southern Medical University. The protocols for animal experimentation were approved by the Southern Medical University Experimental Animal Ethics Committee (no. 00171035 and no. 00181785).
CIA was generated in DBA/1J mice as previously described (35 (link)). Eight‐week‐old male DBA/1J mice were immunized with 100 mg bovine type II collagen (2 mg/ml; Chondrex) emulsified 1:1 in CFA (containing 1 mg/ml M tuberculosis; Chondrex) on day 1 and then injected with Freund's incomplete adjuvant (Chondrex) on day 21.
All animals were maintained under specific pathogen–free conditions in the Laboratory Animal Center of Southern Medical University. The protocols for animal experimentation were approved by the Southern Medical University Experimental Animal Ethics Committee (no. 00171035 and no. 00181785).
6
Collagen-induced Arthritis Model in Mice
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For CIA induction, Female DBA/1 mice were injected subcutaneously at the base of the tail with 100 µg of Chick type II collagen (Condrex, Seattle, WA, USA) emulsified in Freund’s complete adjuvant (CFA) (Chondrex, Seattle, WA, USA), followed 14 days later by a booster injection of the same Chick type II collagen (100 µg) emulsified in Freund’s incomplete adjuvant (Chondrex, Seattle, WA, USA), and mice injected with CFA/IFA only as a negative control. Which via the same route and following the protocol described by David et al (22 (link)). To assess the severity of arthritis, clinical symptoms were evaluated by means of a five-point scale: grade 0 = no swelling; grade 1 = paw with detectable swelling in a single digit; grade 2 = paw with swelling in more than one digit; grade 3 = paw with swelling of all digits and instep; and grade 4 = severe swelling of the paw and ankle.
7
Collagen-Induced Arthritis Rat Model
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CIA animal model was prepared as described by Trentham et al [16 (link)]. The rats were anesthesia via abdominal injection of 35 mg/kg 1% pentobarbital sodium. Then, the 25 mg/ml emulsion was formed by the combination of 500 mg of bovine type II collagen (Chondrex Inc., Washington, U.S.A.) in 1 ml of an emulsion in a 0.3% (Cosmo Bio., Tokyo, Japan) acetic acid solution and 500 mg of Freund’s incomplete adjuvant (Chondrex Inc., Washington, U.S.A.), which was intradermally injected at multiple points on the root of rat’s tail. And the second booster was performed, as 0.5 ml of the same emulsion was intracutaneously injected into the Lt. plantar surface of rats after 3 weeks.
Rats in the CLL group and the CLH group were respectively injected recombinant murine IL-38 intravenously (iv) daily at the base of the tail 1 and 5 ng/g/d for 10 days from the 24th day after the initial immunization, and the control group was injected with the same amount of physiological saline. The skin color, skin temperature, skin infection status of the hind paw and the joints of the rats in each group were observed continuously, and the hind foot movement and the swelling of the joints were observed.
Rats in the CLL group and the CLH group were respectively injected recombinant murine IL-38 intravenously (iv) daily at the base of the tail 1 and 5 ng/g/d for 10 days from the 24th day after the initial immunization, and the control group was injected with the same amount of physiological saline. The skin color, skin temperature, skin infection status of the hind paw and the joints of the rats in each group were observed continuously, and the hind foot movement and the swelling of the joints were observed.
8
Collagen-Induced Arthritis Model
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Bovine type II collagen (Chondrex, Redmond, WA, USA) was diluted to 2 mg/mL with 10 mM acetic acid and emulsified with an equal volume of Freund’s complete adjuvant (Chondrex). The CIA model mice were injected with 0.1 mL of fully emulsified bovine collagen intradermally at the tail base. At 21 days after the first immunization, CIA mice were injected with the same amount of Bovine type II collagen emulsified with Freund’s incomplete adjuvant (Chondrex) as the second immunization.
9
Optimizing Osteoarthritis Treatment with Botanical Compounds
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Bovine type II collagen, Freund's complete adjuvant, and Freund's incomplete adjuvant were purchased from Chondrex Inc. (Washington, USA). Methotrexate (MTX) was purchased from Puri Pharmaceutical (Shanxi, China). Mir-X™ miRNA First Strand Synthesis Kit, Mir-X™ miRNA qRT-PCR SYBR® Kit were purchased from Clontech (Mountain View, USA). Affinity Script QPCR cDNA Synthesis Kit, and Brilliant II SYBR Green QPCR Master Mix Kit were purchased from Agilent (Santa Clara, USA). Human IL-6 was purchased from PEPROTECH. Inc. (Cranbury, USA). The TCM standard Chlorogenic Acid, Loganin, Rutin, Quercetin and Asperosaponin Ⅵ were purchased from National Institutes for Food and Drug Control of China (Beijing, China). HPLC-grade methanol were purchased from Merck (Merck, Darmstadt, Germany). Phosphoric acid purchased in Tianjin Fuyu Fine Chemical Co., Ltd. (Tianjin, China). Water is purchased from Watsons. (Guangdong, China).
10
Collagen-Induced Arthritis Model Protocol
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SKD was prepared according to the previous work [23] (link). Freund's incomplete adjuvant and Bovine Type II Collagen were purchased from Chondrex (Washington DC, USA). ELISA kits of rat TNF-α and IL-1β were ordered from DAKEWE (Shenzhen, China). Raw 264.7 cell line was obtained from National Center for Nanoscience and Technology (Beijing, China). RPMI1640 medium was purchased from Gibco (Carlsbad, USA). Six-well plates were purchased from NEST Biotechnology (Wuxi, China). Other reagents were obtained from commercial vendors.
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