anti type ii collagenmonoclonal antibody cocktail  (Chondrex Inc)

Journal: Scientific Reports

Article Title: Effective CRISPR interference of an endogenous gene via a single transgene in mice

doi: 10.1038/s41598-019-53611-6

Figure Lengend Snippet: Bone phenotype of selected transgenic mouse lines. ( a ) Paraffin sections of the distal femurs of wild type (WT), Tnfsf11 null, and the indicated transgenic mice stained for TRAPase activity. Osteoclasts are stained red. ( b ) Immunohistochemistry of paraffin sections of the distal femurs beneath the growth plate of WT, Tnfsf11 -null, and the indicated transgenic lines for Type II collagen visualization. ( c ) Cancellous bone volume of right femurs from 5-week-old mice from line 157 WT ( n = 6) and transgenic (TG) ( n = 6), line 167 WT ( n = 4) and TG ( n = 5), and line 178 WT ( n = 6) and TG ( n = 3) mice. Both sexes were included. Transgenic compared to WT littermates of the same line using Student’s t test. ( d ) RANKL protein levels in serum of 5-week-old line 157 WT ( n = 7) and TG ( n = 7), line 167 WT ( n = 4) and TG ( n = 5), and line 178 WT ( n = 6) and TG ( n = 3) mice. Both sexes were included. ND, not detected. Transgenic compared to WT littermates of the same line using Student’s t test. ( e ) Osteoprotegerin (OPG) protein levels in serum of 5-week-old line 157 WT ( n = 6) and TG ( n = 7) mice, line 167 WT ( n = 4) and TG ( n = 5), and line 178 WT ( n = 6) and TG ( n = 2). Both sexes were included. Transgenic compared to WT littermates of the same line using Student’s t test. ( f – h ) Quantitative RT-qPCR analysis of Cathepsin K , CalcR , and ACP5 (TRAP) mRNAs in L5 vertebrae from 5-week-old mice of line 157 WT ( n = 6) and TG ( n = 8), line 167 WT ( n = 4) and TG ( n = 5), and line 178 WT ( n = 6) and TG ( n = 3). Both sexes were included. Transgenic compared to WT littermates of the same line using Student’s t test.

Article Snippet: Mouse monoclonal IgG1 COL2A1 antibody (II-II6B3, Developmental Studies Hybridoma Bank, University of Iowa) was diluted to 0.3 μg/mL in PBS with 5% normal horse serum and 1% bovine serum albumin and incubated with sections overnight in a humidified chamber at 4 C. This was followed by washing in PBS-T and incubation with biotinylated anti-mouse IgG for 30 minutes.

Techniques: Transgenic Assay, Staining, Activity Assay, Immunohistochemistry, Quantitative RT-PCR

Journal: International Journal of Molecular Sciences

Article Title: Berberine Delays Onset of Collagen-Induced Arthritis through T Cell Suppression

doi: 10.3390/ijms22073522

Figure Lengend Snippet: The effect of berberine on circulating anti-bovine type II collagen (CII) IgG in the CIA model. ( A ) Anti-CII IgG1, IgG2a, and total IgG at day 28 among all mice (arthritic and non-arthritic) within BBR, PBS (vehicle control), CIA (no treatment control), and non-CIA control animals ( n = 10 per group). ( B ) Anti-CII IgG levels at day 28 compared among arthritic mice only (BBR n = 5; PBS n = 9; CIA n = 9). Statistical comparisons made with the Kruskal–Wallis test with Dunn’s multiple comparisons. ( C ) Anti-CII IgG levels at day 28 compared among BBR-treated mice who developed arthritis (“arthritic”) vs. those that did not (“non-arthritic”). Statistical comparisons made with the Mann–Whitney U test. For all statistical tests in A–C, * p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: Serum concentrations of anti-collagen type II (anti-CII) total IgG (catalog # 1012T), anti-CII IgG1 (catalog # 20321T), and anti-CII IgG2a (catalog # 20322T) were measured by ELISA (Chondrex, Redmond, WA, USA) according to the manufacturer’s instructions [ , ].

Techniques: MANN-WHITNEY

Journal: International Journal of Molecular Sciences

Article Title: Targeting of Phospholipase D1 Ameliorates Collagen-Induced Arthritis via Modulation of Treg and Th17 Cell Imbalance and Suppression of Osteoclastogenesis

doi: 10.3390/ijms21093230

Figure Lengend Snippet: PLD1 deficiency suppresses the collagen type II-specific humoral response and the production of proinflammatory cytokines in collagen-induced arthritis (CIA) mice. ( A ) The amount of anti-collagen total IgG, IgG1, and IgG2a antibodies was measured by ELISA in the serum from the indicated mice at day 42. ( B ) Measurement of proinflammatory cytokines in the serum of the indicated mice at day 42 as analyzed by ELISA. n = 15 per group. * p < 0.05, N.S., non-significant. Results are shown as mean ± standard error of the mean (SEM).

Article Snippet: The amount of murine CII-specific autoantibodies in the serum was measured using the Mouse Anti-Type II Collagen IgG Subtype Antibody Assay Kit (Chondrex), according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Targeting of Phospholipase D1 Ameliorates Collagen-Induced Arthritis via Modulation of Treg and Th17 Cell Imbalance and Suppression of Osteoclastogenesis

doi: 10.3390/ijms21093230

Figure Lengend Snippet: The PLD1 inhibitor reduces the production of the anti-CII IgG2a autoantibody and proinflammatory cytokines in collagen-induced arthritis (CIA) mice. ( A ) The amount of anti-collagen IgG1 and IgG2a antibodies was measured by ELISA in the serum of the indicated mice at day 45. ( B ) The amount of proinflammatory cytokines was measured by ELISA in the serum of the indicated mice at day 45. n = 15 per group. * p < 0.05. Results are shown as mean ± standard error of the mean (SEM).

Article Snippet: The amount of murine CII-specific autoantibodies in the serum was measured using the Mouse Anti-Type II Collagen IgG Subtype Antibody Assay Kit (Chondrex), according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Medicine

Article Title: Humulus japonicus extract ameliorates collagen-induced arthritis in mice through regulation of overall articular inflammation

doi: 10.3892/ijmm.2019.4417

Figure Lengend Snippet: Effect of Humulus japonicas (HJ) on the production of anti-type II collagen antibody in plasma. Levels of (A) anti-type II collagen total IgG and its subtypes (B) IgG1, and (C) IgG2a were measured by ELISA in plasma obtained on day 35 from each mice group. Gene expression levels of (D) Il-12rβ2 , (E) Ccr2 , (F) Ccr5 , (G) Ccr3 , (H) Ccr4 , (I) IL-2 and (J) IL-13 were analyzed by RT-qPCR in the paw of CIA mice on day 35 following the first immunization. The vehicle group was set to a value of 1, and average fold-change is shown. Grouped quantitative data are presented as means ± SEM (vehicle group; n=8, HJ group; n=6). Significance was measured using two-tailed Student's t-test. * P<0.05, ** P<0.01, *** P<0.001.

Article Snippet: Plasma samples were collected at the end of the experiment (day 35) for the determination of IgG, IgG1, and IgG2a antibody levels with three commercially available test kits, mouse anti-mouse Type II collagen IgG TMB (2036T, Chondrex), IgG1 TMB (20361T, Chondrex), and IgG2a (20362T, Chondrex) antibody subtype assay kit TMB, according to the manufacturer's instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Two Tailed Test