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Ab75273

Manufactured by Abcam
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Sourced in Morocco, China

Ab75273 is a laboratory equipment product. It is designed for use in various research applications. The core function of this product is to perform a specific task or procedure in a laboratory setting.

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Lab products found in correlation

Ab51451, by Abcam (1 mentions) Human tgf β1, by R&D Systems (1 mentions) Sc 535, by Santa Cruz Biotechnology (1 mentions) Human fgf9, by R&D Systems (1 mentions) Human bmp4, by R&D Systems (1 mentions) Human bmp6, by R&D Systems (1 mentions) Anti total p44 42 mapk, by Cell Signaling Technology (1 mentions) Anti phospho frs2 α tyr196, by Cell Signaling Technology (1 mentions) Sc 7267, by Santa Cruz Biotechnology (1 mentions) Anti phospho mek1 2, by Cell Signaling Technology (1 mentions) Anti phospho p38, by Cell Signaling Technology (1 mentions) Ab38866, by Abcam (1 mentions) Ab23981, by Abcam (1 mentions) Ab109504, by Abcam (1 mentions) Mouse rabbit igg, by Santa Cruz Biotechnology (1 mentions) Dithiothreitol (dtt), by Roche (1 mentions) Ab5103, by Abcam (1 mentions) Ab1791, by Abcam (1 mentions) Phospho akt antibody, by Cell Signaling Technology (1 mentions) Immobilon psq pvdf membrane, by Merck Group (1 mentions)

5 protocols using ab75273

1

Investigating Intracellular Signaling Pathways

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Recombinant bovine FGF2, human FGF9, human TGFβ1, mouse noggin/Fc chimera, human BMP4, human BMP6, and anti-BMP antibody (MAB3552) were from R&D Systems (Minneapolis, MN). The following antibodies were all purchased from Cell Signaling Technology (Danvers, MA): anti–phospho-p44/42 MAPK E10 mouse monoclonal (#9106), anti–total p44/42 MAPK (#9102), anti–phospho-p38 (#9211), anti–phospho-MEK1/2 (#9154), anti–phospho Raf-1 (#9427), anti–phospho-FRS2-α(Tyr-196) (#3864), and anti–phospho-Smad1 (Ser463/465)/Smad5 (Ser463/465) (#9511). Other antibodies used were as follows: for CP49, rabbit anti-mouse CP49 polyclonal serum (#899 or #900; both generous gifts of Paul FitzGerald, University of California, Davis, CA); for phospho-tyrosine, 4G10 (a kind gift from Brian Druker, Oregon Health and Science University, Portland, OR); for luciferase, #G745A from Promega (Madison, WI); for GFP, JL-8 from Clontech (Mountain View, CA); for phospho-Smad3, ab51451 from Abcam (Cambridge, MA); for total Smad 1/5, ab75273 from Abcam; for total Raf-1, sc-7267 from Santa Cruz Biotechnology (Santa Cruz, CA); for total p38, sc-535 from Santa Cruz; and for total MEK, M17030 from Transduction Labs (Lexington, KT). UO126 (used at 15 μM), PD173074 (100 nM), and dorsomorphin (5 μM) were from Calbiochem (La Jolla, CA). All other reagents, including TPA, were from Sigma-Aldrich (St. Louis, MO).
Boswell B.A, & Musil L.S. (2015). Synergistic interaction between the fibroblast growth factor and bone morphogenetic protein signaling pathways in lens cells. Molecular Biology of the Cell, 26(13), 2561-2572.
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2

Antibody Sourcing for Protein Analysis

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All antibodies were purchased as follows: Antibodies against Smurf1 (ab38866, Abcam), anti-Smad1/5 (ab75273, Abcam), anti-Smad2/3 (#8685, CST), anti-Smad4 (#9515, CST), anti-pSmad1(Ser206) (#13820, CST), anti-Runx2 (ab23981, Abcam), anti-ING2 (ab109504, Abcam), anti-Myc(MBL), anti-GST(MBL), anti-His (MBL), anti-HA (MBL), anti-GAPDH (MBL), anti-Flag (MBL) and mouse/rabbit IgG (Santa Cruz).
Zhang Y., Wang C., Cao Y., Gu Y, & Zhang L. (2016). Selective compounds enhance osteoblastic activity by targeting HECT domain of ubiquitin ligase Smurf1. Oncotarget, 8(31), 50521-50533.
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3

Western Blot Protein Analysis Protocol

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Cells or tissues were lysed on ice in radioimmunoprecipitation assay buffer containing 0.0037 mg/mL protease inhibitor (Roche), 1 mmol/L dithiothreitol (cat. no. D9779; Sigma-Aldrich), 1 mmol/L phenylmethylsulfonyl fluoride (P7626; Sigma-Aldrich), and 100 mmol/L phosphatase inhibitors (P0044 [for cell signaling experiments]; Sigma-Aldrich). Proteins were separated by 10% SDS-PAGE before being transferred onto an Immobilon-PSQ PVDF Membrane (ISEQ-00010; Merck Millipore). Blots were probed with LRG1 antibody (rabbit monoclonal, 13224-1-AP; Proteintech), phosphorylated (phospho-)Smad1/5 antibody (rabbit monoclonal, 9516; Cell Signaling Technology), anti-SMAD1+SMAD5 antibody (mouse monoclonal, ab75273; Abcam), histone H3 (citrulline R2 + R8 + R17) antibody (rabbit polyclonal, ab5103; Abcam), histone H3 antibody (rabbit polyclonal, ab1791; Abcam), phospho-Akt antibody (rabbit monoclonal, 4060; Cell Signaling Technology), Akt antibody (rabbit monoclonal, 9272; Cell Signaling Technology), cyclin D1 antibody (rabbit monoclonal, 2922; Cell Signaling Technology), or GAPDH antibody (rabbit polyclonal, sc-25778; Santa Cruz Biotechnology), followed by horseradish peroxidase–conjugated secondary antibodies (Santa Cruz Biotechnology). Densitometry was performed by use of ImageJ software.
Liu C., Teo M.H., Pek S.L., Wu X., Leong M.L., Tay H.M., Hou H.W., Ruedl C., Moss S.E., Greenwood J., Tavintharan S., Hong W, & Wang X. (2020). A Multifunctional Role of Leucine-Rich α-2-Glycoprotein 1 in Cutaneous Wound Healing Under Normal and Diabetic Conditions. Diabetes, 69(11), 2467-2480.
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4

Immunohistochemical Assessment of Extracellular Matrix Remodeling

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Following antigen retrieval, NP tissue sections were blocked with 5% bovine serum albumin (BSA). Next, the sections were immunostained overnight at 4 °C with the following primary antibodies: MMP13 (rabbit, dilution ratio of 1:500, ab219620, Abcam, Cambridge, UK), Aggrecan (rabbit, dilution ratio of 1:200, ab216965, Abcam), COLII (rabbit, dilution ratio of 1:200, 15943-1-AP, Proteintech, Wuhan, China), and ADAMTS5 (rabbit, dilution ratio of 1:200, PA5-32142, Abcam), p-Smad1/5 (rabbit, dilution ratio of 1:100, ab92698, Abcam), Smad1/5 (mouse, dilution ratio of 1:50, ab75273, Abcam) and Ki67 (rabbit, dilution ratio of 1:200, ab16667, Abcam). Subsequently, the sections were re-probed with biotinylated goat anti-rabbit IgG (dilution ratio of 1:2500, ab205718, Abcam) or goat anti-mouse IgG (dilution ratio of 1:2500, ab6788, Abcam) for 20 min, and then incubated with horseradish peroxidase (HRP)-streptavidin reagent (Sigma, Shanghai, China) for 20 min. Finally, the immunoreactivity was detected by means of DAB staining. Images were obtained under a microscope (Leica-DM2500) and quantified using the ImagePro Plus 7.1 software (Media Cybernetics, Silver Spring, MD, USA).
Yu X.J., Wang Y.G., Lu R., Guo X.Z., Qu Y.K., Wang S.X., Xu H.R., Kang H., You H.B, & Xu Y. (2023). BMP7 ameliorates intervertebral disc degeneration in type 1 diabetic rats by inhibiting pyroptosis of nucleus pulposus cells and NLRP3 inflammasome activity. Molecular Medicine, 29, 30.
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5

Protein Expression in Achilles Tendon Tissues

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Achilles tendon tissues of mice post surgery were obtained and grinded with glass pestles in RIPA buffer on ice before ultrasonication for tissue protein extraction. Cell lysates were collected as described above. Samples were fractionated by 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred to polyvinylidene difluoride membranes (IPVH00010, Millipore). Immunoblotting was performed with primary antibodies against following proteins: Sox9 (1:2000; ab185230, Abcam), Runx2 (1:2000; NBP1-77461, Novusbio), FGFR3 (1:2000; BS90509, bioworld), BMPR1a (1:1000; ab38560, Abcam), pSmad1/5 (1:500; 700047, Thermo), Smad1/5 (1:1000; ab75273, Abcam), β-actin (1:5000; A8481, Sigma-Aldrich). Chemiluminescence was performed with SuperSignal West Dura Extended Duration Substrate (Thermo).
Zhang D., Huang J., Sun X., Chen H., Huang S., Yang J., Du X., Tan Q., Luo F., Zhang R., Zhou S., Jiang W., Ni Z., Wang Z., Jin M., Xu M., Li F., Chen L., Liu M., Su N., Luo X., Yin L., Zhu Y., Feng J.Q., Chen D., Qi H., Chen L, & Xie Y. (2021). Targeting local lymphatics to ameliorate heterotopic ossification via FGFR3-BMPR1a pathway. Nature Communications, 12, 4391.
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