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H 140

Manufactured by Santa Cruz Biotechnology
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Sourced in United States

The H-140 is a laboratory centrifuge designed for general-purpose applications. It features a fixed-angle rotor with a maximum speed of 14,000 RPM and can accommodate sample volumes up to 50 mL. The H-140 is a reliable and versatile instrument suitable for a variety of laboratory procedures that require separation or concentration of samples.

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Lab products found in correlation

Mounting medium, by Agilent Technologies (2 mentions) Sc 393325, by Santa Cruz Biotechnology (2 mentions) Alexa fluor 488 or alexa fluor 594 labelled secondary antibody, by Thermo Fisher Scientific (2 mentions) Ec plan neofluar 40x 1.3 oil, by Zeiss (2 mentions) Diaminiobenzidine as substrate, by Agilent Technologies (2 mentions) Axio imager m1 microscope, by Zeiss (2 mentions) Vectastain elite abc kit, by Vector Laboratories (2 mentions) β catenin antibody, by BD (2 mentions) Hoechst 33342, by Thermo Fisher Scientific (1 mentions) Cm3050s cryostat, by Leica (1 mentions) Mvx10, by Olympus (1 mentions) Ab18184, by Abcam (1 mentions) Odyssey infrared imager 9120, by LI COR (1 mentions) Irdye 800cw goat anti mouse igg secondary antibody, by Abcam (1 mentions) Irdye 800cw goat anti rabbit igg secondary antibody, by LI COR (1 mentions) Horseradish peroxidase conjugated secondary antibody, by Merck Group (1 mentions) Ecl prime, by GE Healthcare (1 mentions)

8 protocols using h 140

1

Cryosectioning and Immunolabeling of Femoral Condyle

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The dissected femoral condyle was immediately embedded in Super Cryo Embedding Medium (Section-lab, Co., Ltd., Hiroshima, Japan) and frozen with dry ice and hexane. Cryosections at 12 μm were prepared with a CM 3050S cryostat (LEICA, Nussloch, Germany) and an ultracut S microtome (Reichert, Vienna, Austria). Rabbit polyclonal anti-lubricin antibody H-140 (1:100 dilution; H-140, Santa Cruz Biotechnology) was applied to sections and incubated at 4°C overnight. Goat anti-mouse IgG labeled with Alexa fluor 555 (1:500 dilution, Invitrogen) was applied and incubated for 1 hour at room temperature. Background nuclei were counterstained with Hoechst 33342 (Invitrogen). Negative controls were incubated without the primary antibody. The specimens were photographed under the condition of fluorescence using Olympus MVX10.
Nakagawa Y., Muneta T., Otabe K., Ozeki N., Mizuno M., Udo M., Saito R., Yanagisawa K., Ichinose S., Koga H., Tsuji K, & Sekiya I. (2016). Cartilage Derived from Bone Marrow Mesenchymal Stem Cells Expresses Lubricin In Vitro and In Vivo. PLoS ONE, 11(2), e0148777.
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2

Immunofluorescence Staining of Tfr2 and Osterix

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For immunofluorescence staining, cells were grown on glass slides. At the desired time point, cells were fixed with 100% methanol for 15 min, permeabilized with 0.5% Triton X-100 for 10 min and after washing for three times, blocked with 1% BSA in PBS for 30 min. Afterwards, cells were incubated with an anti-mouse Tfr2 antibody (H-140, Santa Cruz) over night at 4 °C. After washing, cells were stained with an anti-mouse osterix antibody (sc-393325, Santa Cruz) or phalloidin at RT for 1 h. Subsequently, cells were washed and incubated for 1 h with an Alexa Fluor 488 or Alexa Fluor 594-labelled secondary antibody (Life Technologies), washed, and stained with DAPI for 5 min. After washing again, glass slides were embedded in a small droplet of mounting medium (Dako). Slides were examined using a Zeiss LSM 510 confocal microscope (Zeiss EC Plan-Neofluar 40x/1.3 Oil), and photographs were taken and processed with the Zen 2009 software.
Rauner M., Baschant U., Roetto A., Pellegrino R.M., Rother S., Salbach-Hirsch J., Weidner H., Hintze V., Campbell G., Petzold A., Lemaitre R., Henry I., Bellido T., Theurl I., Altamura S., Colucci S., Muckenthaler M.U., Schett G., Komla Ebri D., Bassett J.H., Williams G.R., Platzbecker U, & Hofbauer L.C. (2019). Transferrin receptor 2 controls bone mass and pathological bone formation via BMP and Wnt signaling. Nature metabolism, 1(1), 111-124.
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3

Immunohistochemical Analysis of Tfr2 and Wnt Signaling

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For immunohistochemical analysis, paraffin sections from WT and Tfr2-/- bones were dewaxed, rehydrated, and heat-retrieved of antigens. Endogenous peroxidase activity was blocked using 0.3% H2O2/PBS for 10 min at room temperature and non-specific binding sites using the blocking buffer of the VECTASTAIN Elite ABC Kit (VECTOR Laboratories) for 45 min at room temperature. Afterwards, sections were incubated with an anti-Tfr2 antibody (H-140, Santa Cruz), a β-catenin antibody (BD Bioscience) or an axin-2 antibody (#ab107613, Abcam) overnight at 4 °C. Subsequently, slides were treated with an anti-mouse secondary antibody conjugated to biotin and then developed utilizing avidin-conjugated HRP with diaminiobenzidine as substrate (DAKO). Slides were examined using a Zeiss Axio Imager M.1 microscope. Two-hundred cells were counted per slide and graded according to no staining (0), weak staining (1), and strong staining (2).
Rauner M., Baschant U., Roetto A., Pellegrino R.M., Rother S., Salbach-Hirsch J., Weidner H., Hintze V., Campbell G., Petzold A., Lemaitre R., Henry I., Bellido T., Theurl I., Altamura S., Colucci S., Muckenthaler M.U., Schett G., Komla Ebri D., Bassett J.H., Williams G.R., Platzbecker U, & Hofbauer L.C. (2019). Transferrin receptor 2 controls bone mass and pathological bone formation via BMP and Wnt signaling. Nature metabolism, 1(1), 111-124.
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4

Immunofluorescence Staining of Tfr2 and Osterix

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For immunofluorescence staining, cells were grown on glass slides. At the desired time point, cells were fixed with 100% methanol for 15 min, permeabilized with 0.5% Triton X-100 for 10 min and after washing for three times, blocked with 1% BSA in PBS for 30 min. Afterwards, cells were incubated with an anti-mouse Tfr2 antibody (H-140, Santa Cruz) over night at 4 °C. After washing, cells were stained with an anti-mouse osterix antibody (sc-393325, Santa Cruz) or phalloidin at RT for 1 h. Subsequently, cells were washed and incubated for 1 h with an Alexa Fluor 488 or Alexa Fluor 594-labelled secondary antibody (Life Technologies), washed, and stained with DAPI for 5 min. After washing again, glass slides were embedded in a small droplet of mounting medium (Dako). Slides were examined using a Zeiss LSM 510 confocal microscope (Zeiss EC Plan-Neofluar 40x/1.3 Oil), and photographs were taken and processed with the Zen 2009 software.
Rauner M., Baschant U., Roetto A., Pellegrino R.M., Rother S., Salbach-Hirsch J., Weidner H., Hintze V., Campbell G., Petzold A., Lemaitre R., Henry I., Bellido T., Theurl I., Altamura S., Colucci S., Muckenthaler M.U., Schett G., Komla Ebri D., Bassett J.H., Williams G.R., Platzbecker U, & Hofbauer L.C. (2019). Transferrin receptor 2 controls bone mass and pathological bone formation via BMP and Wnt signaling. Nature metabolism, 1(1), 111-124.
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5

Western Blot Analysis of Fibrillarin

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Fibrillarin was loaded in 12% acrylamide gel to perform SDS-PAGE, then transferred to a nitrocellulose membrane and blocked with 3% of BSA in PBS at room temperature for 1 h. The membrane then was incubated with rabbit polyclonal anti-Fib antibody (1/3000) (H-140, Santa Cruz Biotechnology, Dallas, TX, USA), for 1 h at room temperature and with the IRDye® 800CW goat anti-rabbit IgG secondary antibody from LICOR for 1 h at room temperature, with three washes of 10 min each with PBS-T between incubations. Immunoblotting signals were analyzed by Odyssey Infrared Imager 9120 (LI-COR Biosciences, Lincoln, NE, USA). For small peptides, the primary antibody was anti-6xHis (1/5000) (Abcam, ab18184, mouse monoclonal) and IRDye® 800CW goat anti-mouse IgG secondary antibody.
Guillen-Chable F., Rodríguez Corona U., Pereira-Santana A., Bayona A., Rodríguez-Zapata L.C., Aquino C., Šebestová L., Vitale N., Hozak P, & Castano E. (2020). Fibrillarin Ribonuclease Activity is Dependent on the GAR Domain and Modulated by Phospholipids. Cells, 9(5), 1143.
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6

Lubricin Expression Profiling

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Conditioned medium for pellet culture between 17 and 21 days and conditioned medium for monolayer culture were collected. 10 μl medium was loaded on each lane separated by electrophoresis with 4–20% gel (Invitrogen). Proteins were transferred onto nitrocellulose membranes by semi wet blotting. Membranes were blocked in 5% skim milk at 4°C overnight and incubated with a rabbit polyclonal anti-lubricin antibody (1:100 dilution; H-140, Santa Cruz Biotechnology, Santa Cruz, CA) at 4°C overnight. Antibodies were detected with horseradish peroxidase-conjugated secondary antibody (Millipore, Billerica, MA) using the enhanced chemiluminescent detection ECL-Prime (GE Healthcare, Little Chalfont, UK).
Nakagawa Y., Muneta T., Otabe K., Ozeki N., Mizuno M., Udo M., Saito R., Yanagisawa K., Ichinose S., Koga H., Tsuji K, & Sekiya I. (2016). Cartilage Derived from Bone Marrow Mesenchymal Stem Cells Expresses Lubricin In Vitro and In Vivo. PLoS ONE, 11(2), e0148777.
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7

Quantitative Analysis of Hes1 Chromatin Binding

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Quantitative ChIP analysis was performed as described previously (29 (link)). Briefly, cells were crosslinked, sonicated, and immunoprecipitated with anti-Hes1 antibodies or IgG (negative control). A mixture of two anti-Hes1 antibodies (H140, Santa Cruz Biotechnology; 4H1, Novus Biologicals) was used for immunoprecipitation. Then, DNA was eluted and crosslinking was reversed, followed by purification and amplification for PCR analysis. The primers used in qPCR are listed in Table III.
Liao B., Zhou H., Liang H, & Li C. (2017). Regulation of ERK and AKT pathways by hepatitis B virus X protein via the Notch1 pathway in hepatocellular carcinoma. International Journal of Oncology, 51(5), 1449-1459.
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8

Immunohistochemical Analysis of Tfr2 and Wnt Signaling

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For immunohistochemical analysis, paraffin sections from WT and Tfr2-/- bones were dewaxed, rehydrated, and heat-retrieved of antigens. Endogenous peroxidase activity was blocked using 0.3% H2O2/PBS for 10 min at room temperature and non-specific binding sites using the blocking buffer of the VECTASTAIN Elite ABC Kit (VECTOR Laboratories) for 45 min at room temperature. Afterwards, sections were incubated with an anti-Tfr2 antibody (H-140, Santa Cruz), a β-catenin antibody (BD Bioscience) or an axin-2 antibody (#ab107613, Abcam) overnight at 4 °C. Subsequently, slides were treated with an anti-mouse secondary antibody conjugated to biotin and then developed utilizing avidin-conjugated HRP with diaminiobenzidine as substrate (DAKO). Slides were examined using a Zeiss Axio Imager M.1 microscope. Two-hundred cells were counted per slide and graded according to no staining (0), weak staining (1), and strong staining (2).
Rauner M., Baschant U., Roetto A., Pellegrino R.M., Rother S., Salbach-Hirsch J., Weidner H., Hintze V., Campbell G., Petzold A., Lemaitre R., Henry I., Bellido T., Theurl I., Altamura S., Colucci S., Muckenthaler M.U., Schett G., Komla Ebri D., Bassett J.H., Williams G.R., Platzbecker U, & Hofbauer L.C. (2019). Transferrin receptor 2 controls bone mass and pathological bone formation via BMP and Wnt signaling. Nature metabolism, 1(1), 111-124.
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