Nb600 450

Manufactured by Novus Biologicals

Sourced in United States

The NB600-450 is a laboratory instrument designed for spectroscopic analysis. It features a wavelength range of 450-600 nm and can be used for various applications that require optical measurements within this spectrum.

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Lab products found in correlation

Eg1160, by Leica (1 mentions) Ab13418, by Abcam (1 mentions) Ab58632, by Abcam (1 mentions) Ab3773, by Abcam (1 mentions) Lsm 780, by Zeiss (1 mentions) Alexa fluor 488 conjugated goat anti mouse igg, by Thermo Fisher Scientific (1 mentions) Normal mouse igg sc 2025, by Santa Cruz Biotechnology (1 mentions) Tween 20, by Merck Group (1 mentions) Hoechst 33258, by Thermo Fisher Scientific (1 mentions) Alexa fluor 555 conjugated goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Bovine serum albumin (bsa), by Fujifilm (1 mentions) Pepsin, by Nacalai Tesque (1 mentions) Rabbit igg sc 2027, by Santa Cruz Biotechnology (1 mentions) Triton x, by Fujifilm (1 mentions) Fl 95, by Santa Cruz Biotechnology (1 mentions) Sp1600, by Leica (1 mentions) Sc 393859, by Santa Cruz Biotechnology (1 mentions) Nbp1 05119, by Novus Biologicals (1 mentions) Nb7539, by Novus Biologicals (1 mentions) Sc 365797, by Santa Cruz Biotechnology (1 mentions)

Spelling variants of this product

COL1A1 (NB600–450), (2 citations)

6 protocols using nb600 450

Histological and Immunohistochemical Evaluation of Osteochondral Defects

Cited 2 times

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After the micro-CT analysis, the samples were decalcified in 15% (w/v) ethylenediamine-tetraacetic acid (EDTA), dehydrated through series of ethanol, embedded in paraffin, and longitudinally sectioned into slices with an approximate thickness of 5 μm using a paraffin microtome (Leica EG 1160). The sections were subsequently stained with hematoxylin and eosin (H&E), safranin-fast green (S-F), toluidine blue (T-B) and Masson’s trichrome (M-T) to observe new tissues. The histological sections were blindly and independently scored by three evaluators, using an established histological scoring system (Table S2) for osteochondral defects [38 (link)–40 (link)]. The scores were averaged to determine the final scores.
For immunohistochemical (IHC) staining of collagen type I (COL I), collagen type II (COL II), collagen type X (COL X), aggrecan (AGG) and osteocalcin (OCN), sections were immersed in 0.3% (w/v) H₂O₂, and blocked in 5% (v/v) BSA solution. Followed by enzymatic antigen retrieval, the sections were incubated with primary antibodies against COL I (NB600-450, Novus), COL II (NBP2-33343, Novus), COL X (ab58632, Abcam), AGG (ab3773, Abcam), and OCN (ab13418, Abcam). After rinsing with PBS, they were incubated with horseradish peroxidase-conjugated IgG, followed by 3, 3-diaminobenzidiine tetrahydrochloride (DAB) for visualization. Nuclei were counterstained with hematoxylin.

Du Y., Liu H., Yang Q., Wang S., Wang J., Ma J., Noh I., Mikos A.G, & Zhang S. (2017). Selective Laser Sintering Scaffold with Hierarchical Architecture and Gradient Composition for Osteochondral Repair in Rabbits. Biomaterials, 137, 37-48.

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Immunofluorescent Analysis of Bone Matrix Proteins

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Antigen retrieval was performed via incubation of specimens in a solution of 0.5 M acetic acid (Sigma-Aldrich) with 0.1% pepsin (Nacalai Tesque) at 37°C for 1 hour in a humid chamber prior to immunofluorescent staining. After washing, nonspecific binding was blocked for 60 minutes using 2% BSA (Wako Pure Chemical), 0.1% Tween 20 (Sigma-Aldrich), and 0.01% Triton-X (Wako Pure Chemical) prior to incubation in the primary antibody at 4°C overnight. After washing, the specimens were incubated with the secondary antibody for 1 hour at room temperature. Subsequently, fluorescence was detected by confocal microscopy (LSM780, Zeiss). The primary antibodies used in this study were anti-type I collagen monoclonal antibody (NB600-450: 1/50, Novus Biologicals, Littleton, CO, USA), anti-osteocalcin polyclonal antibody (FL-95: 1/50, Santa Cruz Biotechnology), and control IgG (normal mouse IgG (sc-2025): 1/50 or rabbit IgG (sc-2027): 1/50, Santa Cruz Biotechnology). Secondary antibodies were Alexa Fluor 488-conjugated goat anti-mouse IgG (1/500, Molecular Probes, Thermo Fisher Scientific) or Alexa Fluor 555-conjugated goat anti-rabbit IgG (1/500, Thermo Fisher Scientific). Nuclear staining was performed using Hoechst 33258 (1/500, Thermo Fisher Scientific).

Limraksasin P., Okawa H., Zhang M., Kondo T., Osathanon T., Pavasant P, & Egusa H. (2020). Size-Optimized Microspace Culture Facilitates Differentiation of Mouse Induced Pluripotent Stem Cells into Osteoid-Rich Bone Constructs. Stem Cells International, 2020, 7082679.

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Evaluating Femur Cartilage Structure

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Five femur condyles in each group were fixed in 10% formalin for 2 days, then decalcified using 10% EDTA solution (pH 7.4) for 21 days, finally embedded in paraffin. 5 μm sections of each specimen were collected. Hematoxylin and eosin (H&E) staining was implemented to examine the morphology.
After the deparaffinized sections were prepared, Immunohistochemistry was performed as previously describe53 (link). Briefly, the sections were incubated with primary collagen I antibody ((NB600-450, Novus Biologicals, USA) and osteopontin (OPN) antibody (clone 1B20; Novus Biologicals, USA) at 4 °C for overnight, and then incubated with goat anti-mouse biotin-conjugated IgG (Vector Lab Inc, USA) for 1 h. Subsequently, color reaction was developed with diaminobenzidine (Dako, USA). Finally, the sections were counterstained with hematoxylin and examined under a Leica microscope.
The other five femur condyles in each group were fixed in acetone for 7 days, dehydrated in graded series of alcohol for 3 days each, and embedded in methyl methacrylate without decalcification. The non-decalcified sections, which were parallel to the long axis of the femur, were made on a diamond saw (Leica SP1600) and grounded to about 50 μm in thickness. Finally, the specimens were stained with Van Gieson’s picrofuchsin staining.

Cheng M.Q., Wahafu T., Jiang G.F., Liu W., Qiao Y.Q., Peng X.C., Cheng T., Zhang X.L., He G, & Liu X.Y. (2016). A novel open-porous magnesium scaffold with controllable microstructures and properties for bone regeneration. Scientific Reports, 6, 24134.

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Immunohistochemical Analysis of Extracellular Matrix Proteins

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Immunohistochemistry was performed for collagen type 1 (dilution, 1:100; collagen 1 alpha 1 antibody; catalog No. NB600–450; Novus Biologicals), collagen type 3 (dilution, 1:100; collagen 3 alpha 1 antibody; catalog No. NBP1–05119; Novus Biologicals), tenomodulin (dilution, 1:50; tenomodulin antibody aa74–123; Horseradish Peroxidase (HRP) conjugated; catalog No. LS-C452705; LifeSpan BioSciences), and matrix metalloproteinase 9 (MMP-9) (dilution, 1:50; catalog No. sc-393859; Santa Cruz Biotechnology). A conjugated secondary antibody (dilution, 1:500; Goat anti-Mouse Immunoglobulin G (IgG) HRP; catalog No. HAF007; Novus Biologicals) was utilized with unconjugated primary antibodies (collagen 1, collagen 3, and MMP-9).

McClellan P., Ina J.G., Knapik D.M., Isali I., Learn G., Valente A., Wen Y., Wen R., Anderson J.M., Gillespie R.J, & Akkus O. (2022). Mesenchymal Stem Cell Delivery via Topographically Tenoinductive Collagen Biotextile Enhances Regeneration of Segmental Tendon Defects. The American journal of sports medicine, 50(8), 2281-2291.

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Histological Evaluation of Tissue Samples

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For histological examination, samples were fixed in 4% paraformaldehyde for 2 days, embedded in paraffin, and sectioned into a series of 6 μm slices. The sections were further stained with hematoxylin and eosin (H&E), toluidine blue (TB), and Sirius red (SR) according to the manufacturer's protocols. Additionally, the sections were treated for antigen retrieval and incubated with primary antibodies against collagen I (1:100, Cat. No. NB600‐450; Novus) and collagen II (1:100, Cat. No. NBP2‐33343) overnight at 4°C, followed by incubation with a goat anti‐mouse IgG secondary antibody (1:200; Cat# NB7539; Novus) for 1 h at room temperature. The diaminobenzidine (DAB) substrate system was used for color development.

Li H., Zhao T., Cao F., Deng H., He S., Li J., Liu S., Yang Z., Yuan Z, & Guo Q. (2022). Integrated bioactive scaffold with aptamer‐targeted stem cell recruitment and growth factor‐induced pro‐differentiation effects for anisotropic meniscal regeneration. Bioengineering & Translational Medicine, 7(3), e10302.

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Histological Evaluation of Bone Regeneration

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Hematoxylin and eosin (H&E) stain, Masson’s trichrome stain, and immunohistochemical (IHC) stain tests were used to study new bone histology and were performed after the harvested calvarial samples were subjected to micro CT scanning. The samples fixed with 10% formalin were again decalcified using ethylene-diamine-tetraacetic acid solution. After demineralization, the samples were embedded in paraffin and sliced into samples with 4 μm thicknesses. Afterward, the slides were stained with hematoxylin and eosin (H&E) and Masson’s trichrome stain. Masson’s trichrome stain was performed in accordance with the manual instruction of Masson’s trichrome stain kit (ab150686, Abcam, Cambridge, UK). The other slides were immunohistochemically stained for type I collagen (NB600-450, Novus Biologicals, Littleton, CO, USA) and osteocalcin (sc-365797, Santa Cruz Biotechnology, Santa Cruz, CA, USA) to evaluate new bone formation characteristics. In order to analyze the trends in collagen type I and osteocalcin protein staining from the IHC-stained images, the stained area relative to the total tissue area, excluding the scaffold area, was calculated using the Image J (Version 1.53q; National Institutes of Health, Bethesda, MD, USA) analysis tool.

Kim Y.M., Ghim M.S., Quan M., Kim Y.Y, & Cho Y.S. (2024). Experimental Verification of the Impact of the Contact Area between the Defect Site and the Scaffold on Bone Regeneration Efficacy. Polymers, 16(3), 338.

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