Rabbit anti osteopontin

Manufactured by Abcam

Sourced in Czechia, United States

Rabbit anti-osteopontin is a primary antibody that detects osteopontin, a protein involved in various cellular processes. It can be used for research applications such as Western blotting, immunohistochemistry, and immunofluorescence.

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Lab products found in correlation

Triton x 100, by Merck Group (1 mentions) Rabbit anti ki67 antibody, by Abcam (1 mentions) Paraformaldehyde (pfa), by Merck Group (1 mentions) Bovine serum albumin faction 5, by Roche (1 mentions) Rabbit anti collagen 1, by Abcam (1 mentions) Lsm 510 meta confocal laser scanning microscope, by Zeiss (1 mentions) Mouse anti β 3 tubulin, by Merck Group (1 mentions) Alexa 488 phalloidin, by Thermo Fisher Scientific (1 mentions) Glass coverslip, by Thermo Fisher Scientific (1 mentions) Human msc, by Lonza (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Cell culture media, by Thermo Fisher Scientific (1 mentions) Tetramethylrhodamine conjugated anti rabbit igg antibody, by Thermo Fisher Scientific (1 mentions) Chicken anti map2, by Abcam (1 mentions) Ab5392, by Abcam (1 mentions) Ab23981, by Abcam (1 mentions) Ab8448, by Abcam (1 mentions) Rabbit anti runx2, by Abcam (1 mentions) T8660, by Merck Group (1 mentions) Multi analyte elisarray kit, by Qiagen (1 mentions)

Close spelling variants of this product

Osteopontin (25 citations) Anti-osteopontin (18 citations) Rabbit anti- (5 citations) Anti-rabbit osteopontin primary antibody (3 citations)

4 protocols using rabbit anti osteopontin

Immunostaining of OCCM-30 Cells

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OCCM-30 cells were seeded on glass coverslips (Thermanox) and treated as described above. After one and two days, cells were fixed with 4% paraformaldehyde (Merck KGaA, Darmstadt, Germany) for 10 min and permeabilized with 0.05% Triton X-100 (Merck) in PBS. Blocking of non-specific binding was performed with 5% bovine serum albumin faction V (Roche Diagnostics, Indianapolis, IN, USA) in 1× PBS for 45 min at room temperature. Primary antibodies were incubated for 1,5 h at room temperature or overnight at 4 °C: rabbit polyconal anti-collagen I antibody (1:200; Abcam, Cambridge, UK), rabbit polyclonal anti-osteoblast specific factor 2 antibody (1:1000; BioVendor, Brno, Czech Republic), rabbit anti-osteopontin (1:200; Abcam), and rabbit anti-Ki67 antibody (1:200; Abcam). Peroxidase-conjugated anti-mouse or anti-rabbit EnVision^® (Dako, Glostrup, Denmark) were used as secondary antibodies for 30 min at room temperature and visualized with diaminobenzidine (DAB). OCCM-30 cells were counterstained with hemalaunum (Merck). Semi-quantitative analysis of stained cells was performed using ImageJ Fiji (ImageJ 2.1.0/1.53c, National Institutes of Health, Bethesda, MD, USA) as described in the literature [79 (link)]. In each case, 3 sections of 3 independent experiments were randomly selected for measurement.

Eggers B., Marciniak J., Deschner J., Stope M.B., Mustea A., Kramer F.J, & Nokhbehsaim M. (2021). Cold Atmospheric Plasma Promotes Regeneration-Associated Cell Functions of Murine Cementoblasts In Vitro. International Journal of Molecular Sciences, 22(10), 5280.

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Immunofluorescent Characterization of BMSC-Seeded Scaffolds

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At day 14 and day 28, scaffold samples seeded with BMSCs were collected and prepared for immunofluorescent staining and microscopy. Primary antibodies included rabbit anti-Collagen 1 (dilution 1:500), rabbit anti-osteopontin (dilution 1:500) (Abcam, Cambridge, MA). Cells were fixed in 4% paraformaldehyde solution pH 6.9 for 20 min at room temperature, followed by three PBS rinses. Samples were permeabilized with 0.2% Triton X-100 for 5 min at room temperature. Following PBS rinses, samples were blocked using 5% normal goat serum for 1 h at room temperature. In separate experiments, primary antibodies of the aforementioned protein markers were diluted appropriately in blocking solution and samples were allowed to incubate overnight at 4 °C. Post PBS rinses, fluorescent-labeled secondary antibody (antirabbit IgG; Abcam, Cambridge, MA) were applied for 1 h at room temperature in the dark. Following incubation, the spiral structures were viewed using a Zeiss LSM 510 Meta confocal laser scanning microscope (Carl Zeiss, Oberkochen, Germany).

Manoukian O.S., Aravamudhan A., Lee P., Arul M.R., Yu X., Rudraiah S, & Kumbar S.G. (2018). Spiral Layer-by-Layer Micro-Nanostructured Scaffolds for Bone Tissue Engineering. ACS biomaterials science & engineering, 4(6), 2181-2192.

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Immunofluorescence Microscopy of Differentiated Cells

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Laboratory chemicals and reagents were purchased from Sigma Aldrich unless otherwise noted. Tissue culture plastic ware was purchased from Thermo Fisher Scientific. Cell culture media and reagents were purchased from Gibco. Human MSCs and differentiation media were purchased from Lonza. Rabbit anti-Runx2 was purchased from abcam (ab23981) Technologies, rabbit anti-Osteopontin was purchased from abcam (ab8448), mouse anti-β3 tubulin was purchased from Sigma (T8660), and chicken anti-MAP2 was purchased from abcam (ab5392). Tetramethylrhodamine-conjugated anti-rabbit IgG antibody, Alexa488-phalloidin, Alexa647-conjugated anti-mouse IgG, Alexa647-conjugated anti-chicken IgG, and 4′,6-diamidino-2-phenylindole (DAPI) were purchased from Invitrogen. Glass coverslips (18-mm circular) for surface preparation were purchased from Fisher Scientific.

Lee J., Abdeen A.A, & Kilian K.A. (2014). Rewiring mesenchymal stem cell lineage specification by switching the biophysical microenvironment. Scientific Reports, 4, 5188.

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Immune Markers in Bone Marrow Mutants

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Equal amounts (20 µg/sample) of protein extract obtained from BM cells of mutants and WT littermates were run on 10% to 12% SDS-PAGE followed by blotting onto polyvinyl difluoride membranes. Blots were probed with primary antibodies specific to rabbit-anti-polyclonal SDF-1 (cat.#sc-28876, 1:200; Santa Cruz Biotechnology), rabbit-anti-osteopontin (cat.#ab8448; Abcam), rabbit-anti-runt-related transcription factor 2 (Runx2) (cat.#BS2831; Bioworld Technology, USA), rabbit-anti-osterix (cat.#ab94744; Abcam), or mouse-anti-monoclonal β-actin (cat.#sc-81178, 1:200; Santa Cruz Biotechnology) at 4°C. Membranes were washed and exposed to horseradish peroxidase-conjugated rabbit-anti IgG or mouse-anti Ig. Immunoreactive bands were visualized by enhanced chemiluminescence (Santa Cruz Biotechnology) before exposure to X-ray film (Eastman Kodak, USA). To evaluate the level of SDF-1 in BM supernatants or cultured BM cells, ELISA was performed using an SDF-1 mouse ELISA kit (Abcam) following the manufacturers’ instructions. The levels of inflammatory cytokines such as interleukin (IL)-1α, IL-6, interferon (IFN)-γ, and tumor necrosis factor (TNF)-α in BM supernatants of mutants and WT littermates were measured using Multi-Analyte ELISArray Kits according to the manufacturer’s instructions (Qiagen Sciences, Germany).

Sim H.J., Kim M.H., Bhattarai G., Hwang J.W., So H.S., Poudel S.B., Cho E.S., Kook S.H, & Lee J.C. (2021). Overexpression of COMP-Angiopoietin-1 in K14-Expressing Cells Impairs Hematopoiesis and Disturbs Erythrocyte Maturation. Molecules and Cells, 44(4), 254-266.

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