mouse polyclonal anti aggrecan  (Bioss)

Journal: Journal of Musculoskeletal & Neuronal Interactions

Article Title: Changes in growth plate extracellular matrix composition and biomechanics following in vitro static versus dynamic mechanical modulation

doi:

Figure Lengend Snippet: A) Representative aggrecan immunohistochemical slides; B) Average aggrecan reaction signal intensity. Values are expressed as mean ± SD. Significant differences are marked with * (p<0.01). BL, baseline; Ctrl, control; Stat, static; Dyn1, dynamic at 30% amplitude, 0.1 Hz frequency, Dyn2, dynamic at 30% amplitude, 1.0 Hz frequency; Dyn3, dynamic at 100% amplitude, 0.1 Hz frequency; IHC, Immunohistochemistry.

Article Snippet: The sections were incubated overnight at 4°C in presence of primary antibody: polyclonal rabbit anti-mouse anti-aggrecan (1/50; Chemicon, Temecula, CA), monoclonal mouse anti-chicken anti-type II collagen (1/50; DSHB, Iowa, USA) or monoclonal mouse anti-porcine anti-type X collagen (1/50; Sigma-Aldrich).

Techniques: Immunohistochemical staining, Immunohistochemistry

Journal: Regeneration

Article Title: Analogous cellular contribution and healing mechanisms following digit amputation and phalangeal fracture in mice

doi: 10.1002/reg2.51

Figure Lengend Snippet: P2 amputation initiates local endochondral ossification response. (A) Mallory trichrome stained section of an adult mouse hind limb digit. Dashed line indicates the amputation level. (B) μCT reconstructed images show new bone formation initiated by 14 DPA, and continued bone formation at 21 and 28 DPA. (C) Bone volume measurements, normalized to 1 DPA, indicate bone degradation at 7 DPA, resulting in an average bone loss of 12%, followed by a bone growth phase, resulting in an 18% overall increase in bone volume by 28 DPA ( t test, ±SEM, * P < 0.005). (D)−(H) Histological sections of digits stained with Mallory trichrome stain. (D), (D′) At 6 DPA wound closure has not completed and wound retraction is evident. (E), (E′) Wound closure is completed by 9 DPA, and cartilaginous growth is evident on the lateral portions of the bone (outlined). (F), (F′) Replacement of the cartilaginous callus with woven bone and marrow cells is apparent by 15 DPA. (G), (G′) By 24 DPA, a bone plug has formed at the distal portion of the stump. (H) Bone remodeling and tendon reattachment to the bone (arrowhead) is evident by 45 DPA. (I)−(K) Immunostaining of 9 DPA samples, counterstained with DAPI. (I) Col II immunostaining indicates cartilaginous tissue along the periosteal surface. (J) Immunostaining for aggrecan (ACAN) confirming cartilaginous tissue is present. (K) Immunostaining for osterix (Osx) reveals osteoblasts localized to the periosteal callus and the endosteum. (A), (D)−(K) Dorsal surface is top, distal to the right. (B) Top is proximal, bottom is distal. S, scab; WE, wound epidermis; RBC, red blood cells. Scale bars: (A) 500 μm; (D)−(H) 200 μm; (I)−(K) 50 μm.

Article Snippet: Immunostaining for aggrecan was performed using the rabbit anti‐mouse aggrecan polyclonal antibody (EMD Millipore, Billerica, MA) and the Alexa Fluor 488 goat anti‐rabbit IgG secondary antibody (Invitrogen).

Techniques: Staining, Immunostaining

Journal: Journal of Zhejiang University. Science. B

Article Title: Adenovirus-mediated GDF-5 promotes the extracellular matrix expression in degenerative nucleus pulposus cells *

doi: 10.1631/jzus.B1500182

Figure Lengend Snippet: Primer sequences used for RT-PCR

Article Snippet: The cells were then developed with a brown specific stain by 3,3'-diaminobenzidine (DAB) substrate solution for 30 min. For visualization and aggrecan assay, the cells were treated as above, blocked with goat IgG (1:250, v/v) for 2 h at room temperature and then incubated with rabbit anti-mouse aggrecan polyclonal antibody (5 mg/ml; Santa Cruz Biotechnology, Santa Cruz, CA, USA) for 12–18 h at 4 °C, followed by incubation with fluorescein isothiocyanate (FITC; for aggrecan) conjugated secondary antibody for 1 h at room temperature in a moist dark box.

Techniques:

Journal: Journal of Zhejiang University. Science. B

Article Title: Adenovirus-mediated GDF-5 promotes the extracellular matrix expression in degenerative nucleus pulposus cells *

doi: 10.1631/jzus.B1500182

Figure Lengend Snippet: Immunostaining of human NP cells for type-II collagen and aggrecan after they were infected by Ad-GDF-5, Ad-GFP, or no adenovirus (BM group)

Article Snippet: The cells were then developed with a brown specific stain by 3,3'-diaminobenzidine (DAB) substrate solution for 30 min. For visualization and aggrecan assay, the cells were treated as above, blocked with goat IgG (1:250, v/v) for 2 h at room temperature and then incubated with rabbit anti-mouse aggrecan polyclonal antibody (5 mg/ml; Santa Cruz Biotechnology, Santa Cruz, CA, USA) for 12–18 h at 4 °C, followed by incubation with fluorescein isothiocyanate (FITC; for aggrecan) conjugated secondary antibody for 1 h at room temperature in a moist dark box.

Techniques: Immunostaining, Infection

Journal: Journal of Zhejiang University. Science. B

Article Title: Adenovirus-mediated GDF-5 promotes the extracellular matrix expression in degenerative nucleus pulposus cells *

doi: 10.1631/jzus.B1500182

Figure Lengend Snippet: Concentrations of collagen II (Col II) and aggrecan in NP cells at 21th days post-infection by ELISA

Article Snippet: The cells were then developed with a brown specific stain by 3,3'-diaminobenzidine (DAB) substrate solution for 30 min. For visualization and aggrecan assay, the cells were treated as above, blocked with goat IgG (1:250, v/v) for 2 h at room temperature and then incubated with rabbit anti-mouse aggrecan polyclonal antibody (5 mg/ml; Santa Cruz Biotechnology, Santa Cruz, CA, USA) for 12–18 h at 4 °C, followed by incubation with fluorescein isothiocyanate (FITC; for aggrecan) conjugated secondary antibody for 1 h at room temperature in a moist dark box.

Techniques: Infection, Enzyme-linked Immunosorbent Assay