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Ertr7

Manufactured by Santa Cruz Biotechnology
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ERTR7 is a laboratory reagent used for the detection and analysis of specific cellular markers in biological samples. It functions as an antibody that can bind to and identify the presence of a particular target protein or antigen within cells. The core function of ERTR7 is to provide researchers with a tool for cellular characterization and identification purposes.

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Lab products found in correlation

Keratin17, by Cell Signaling Technology (2 mentions) Tgf βrii, by Santa Cruz Biotechnology (1 mentions) F4 80, by Santa Cruz Biotechnology (1 mentions) Il 11rα1, by Santa Cruz Biotechnology (1 mentions) Sftpc, by Abcam (1 mentions) Il 11, by Santa Cruz Biotechnology (1 mentions) Tgf β1, by Abcam (1 mentions) Vimentin, by Cell Signaling Technology (1 mentions) Collagenase 1, by Thermo Fisher Scientific (1 mentions) Anti cxcl13, by R&D Systems (1 mentions) Hydrocortisone, by Thermo Fisher Scientific (1 mentions) Human epidermal growth factor (hegf), by Thermo Fisher Scientific (1 mentions) Foetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Medium 254, by Thermo Fisher Scientific (1 mentions) Heparin, by Thermo Fisher Scientific (1 mentions) Phorbol 12 myristate 13 acetate, by Thermo Fisher Scientific (1 mentions) Epilife medium, by Thermo Fisher Scientific (1 mentions) Basic fibroblast growth factor (bfgf), by Thermo Fisher Scientific (1 mentions)

7 protocols using ertr7

1

Multimarker Immunofluorescence Staining

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Primary antibodies against CD3 (sc-20047, Santa Cruz Biotechnology Inc.), F4/80 (sc-377009, Santa Cruz Biotechnology Inc.), IL-11Rα1 (sc-130920, Santa Cruz Biotechnology Inc.), SFTPC (ab211326, Abcam), TGF-β1 (ab64715, Abcam), IL-11 (sc-133063, Santa Cruz Biotechnology Inc.), vimentin (#5741, Cell Signaling Technology), p53 (#2524, Cell Signaling Technology), p16 (#MA5-17142, Invitrogen Inc.), fibroblast marker (ER-TR7) (sc-73355, Santa Cruz Biotechnology Inc.), and TGF-β RII (sc-17792, Santa Cruz Biotechnology Inc.) and affinity-purified Alexa Fluor 488-conjugated and 594-conjugated secondary antibodies (Life Technologies Corporation, USA) were used. Details are provided in Supporting Information 3.
Chen H., Chen H., Liang J., Gu X., Zhou J., Xie C., Lv X., Wang R., Li Q., Mao Z., Sun H., Zuo G., Miao D, & Jin J. (2020). TGF-β1/IL-11/MEK/ERK signaling mediates senescence-associated pulmonary fibrosis in a stress-induced premature senescence model of Bmi-1 deficiency. Experimental & Molecular Medicine, 52(1), 130-151.
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2

Isolation and Culture of Mouse Prostate Fibroblasts

Cited 1 time
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Prostates were collected from six to eight-week-old C57BL/6 mice [12 (link)]. After mincing the prostates, they were digested by 5 mg/mL of collagenase I (Thermo Fisher Scientific, Waltham, MA, USA, cat. 17100017) for 30 min at 37 °C [23 (link)]. Tissue fragments were then cultured for 2–3 weeks in Dulbecco’s Minimal Essential Media (DMEM, Hyclone, Logan, UT, USA) supplemented with 10% fetal bovine serum (FBS), 1% penicillin/streptomycin and 1% non-essential amino acids. After 5 days of culture, all the cells were fibroblasts. The purity of the cells were determined by ERTR7 (Santa Cruz, cat. sc-73355), a fibroblast marker and Keratin17 (Cell Signaling cat. 4543) an epithelial cell marker. All experiments were repeated in triplicate using primary fibroblasts cells collected from prostates of different mice.
Chatterjee A., Zhu Y., Tong Q., Kosmacek E.A., Lichter E.Z, & Oberley-Deegan R.E. (2018). The Addition of Manganese Porphyrins during Radiation Inhibits Prostate Cancer Growth and Simultaneously Protects Normal Prostate Tissue from Radiation Damage. Antioxidants, 7(1), 21.
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3

Tissue Preparation and Immunofluorescence Staining

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CCL2-GFP tissues were fixed in 4% paraformaldehyde (1 hour on ice), cryoprotected in 30% sucrose, and frozen in optimal cutting temperature (OCT) embedding medium (Tissue-Tek, Torrance, CA). Other tissues were fresh-frozen in OCT. Antibodies are as used for FACS except anti-CXCL13 (R&D Systems), ERTR7 (Santa Cruz Biotechnologies, Santa Cruz, CA), GFP-Alexa488 and FITC-Alexa488 (both Thermo Fisher Scientific), and goat-Alexa488, rat-rhodamine, armenian hamster-AMCA, rabbit-rhodamine, mouse IgG-biotin, and streptavidin-rhodamine/AMCA (all Jackson Immunoresearch).
Dasoveanu D.C., Park H.J., Ly C.L., Shipman W.D., Chyou S., Kumar V., Tarlinton D., Ludewig B., Mehrara B.J, & Lu T.T. (2020). Lymph node stromal CCL2 limits antibody responses. Science immunology, 5(45), eaaw0693.
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4

Isolation and Culture of Skin Cell Types

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Adult skin specimens obtained from Caesarean sections and circumcisions were used to establish the cell cultures. Keratinocyte cultures were obtained by suspending individual epidermal cells derived according to published methods (22 (link)), in EpiLife Medium supplemented with bovine pituitary extract (BPE), bovine insulin (BI), hydrocortisone, human epidermal growth factor, and bovine transferrin (BT) (Invitrogen). Early passages from 3 to 4 were used for these experiments. Melanocyte cultures were obtained by suspending individual epidermal cells in Medium 254 supplemented with BPE, foetal bovine serum (FBS), BI, hydrocortisone, bFGF, BT, heparin, and phorbol 12-myristate 13-acetate (Invitrogen). Fibroblast cultures were obtained by suspending individual dermal cells lacking epidermis (23 ) in DMEM-supplemented with 10% FBS, 100 U/ml penicillin, and 0.1 mg/ml streptomycin (Invitrogen). Cytokeratins 14 and 10 (mouse monoclonal; Santa Cruz Biotechnology, Dallas, TX, USA) were used as markers for keratinocytes; c-kit (mouse monoclonal; Santa Cruz Biotechnology) and tyrosinase-related protein 1 (goat polyclonal; Santa Cruz Biotechnology) were used as markers for melanocytes; and ER-TR7 (rat polyclonal; Santa Cruz Biotechnology) as a marker for fibroblasts (24 (link)–26 (link)).
CHEONG K.A, & LEE A.Y. (2020). Guanine Deaminase Stimulates Ultraviolet-induced Keratinocyte Senescence in Seborrhoeic Keratosis via Guanine Metabolites. Acta Dermato-Venereologica, 100(8), 5732.
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5

Immunohistochemical Analysis of MMP-12 and TIMP-2

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Paraffin‐embedded tissue sections were deparaffinized, and antigens were retrieved for 5 min in a pressure cooker at 121°C in pH 9.0 antigen retrieval solution (Nichirei Bioscience). For MMP‐12 staining, the sections were incubated with a rabbit monoclonal antibody (1:100 dilution, BS9869M, Bioworld Technology) at room temperature for 60 min. Sites of antibody binding were visualized with the Histofine simple stain MAX‐PO(R) kit (Nichirei Bioscience). 3,3′‐Diaminobenzidine tetrahydrochloride was used as a chromogen, and the sections were counterstained with hematoxylin. For TIMP‐2 staining, a mouse monoclonal antibody (1:100 dilution, 3A4, Santa Cruz Biotechnology) was applied at room temperature for 60 min and visualized with the Histofine mouse staining kit (Nichirei Bioscience). For immunofluorescence, a rabbit polyclonal antibody against p19ARF (1:300 dilution, ab80, Abcam) and a rat monoclonal antibody against fibroblasts (1:50 dilution, ER‐TR7, Santa Cruz Biotechnology) were used. The sections were visualized with Alexa Fluor 488‐conjugated anti‐rat IgG and Alexa594‐conjugated anti‐rabbit IgG and were counterstained with DAPI.
Mikawa R., Suzuki Y., Baskoro H., Kanayama K., Sugimoto K., Sato T, & Sugimoto M. (2018). Elimination of p19ARF‐expressing cells protects against pulmonary emphysema in mice. Aging Cell, 17(5), e12827.
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6

Isolation and Culture of Primary Mouse Colonic Fibroblasts

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Colons/rectums were isolated from 8-10 week old C57BL/6 mice and digested in 5 mg/mL collagenase I [54 ]. Cells and large tissue fragments were divided and cultured for 2-3 weeks in Dulbecco's Minimal Essential Media (DMEM) supplemented with 10% fetal bovine serum (FBS), 1% penicillin/streptomyocin and 1% non-essential amino acids with or without 0.25 μM MnTnBuOE-2-PyP. Media and MnTnBuOE-2-PyP were replaced every three days or the cells were passaged at confluency. Cell types in the cultures were characterized using immunofluorescence for Keratin17 (Cell Signaling #4543) to detect contaminating epithelial cells and ERTR7 (Santa Cruz #sc-73355) to verify the presence of fibroblasts. Epithelial cells were not detected in any of the cultures used for experimentation. All experiments were performed in the second or third week of culturing when cell division and viability was maximal. All assays were repeated in triplicate using separate isolations of primary fibroblasts.
Kosmacek E.A., Chatterjee A., Tong Q., Lin C, & Oberley R.E. (2016). MnTnBuOE-2-PyP protects normal colorectal fibroblasts from radiation damage and simultaneously enhances radio/chemotherapeutic killing of colorectal cancer cells. Oncotarget, 7(23), 34532-34545.
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7

Quantifying Apoptosis in Lymph Nodes

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Sections from frozen LNs were fixed in 4% paraformaldehyde, stained with Masson Trichrome (Sigma) to detect collagen deposition according to manufacturer’s instructions; Primary antibodies used for immunofluorescence staining: ERTR7 (Santacruz; sc73355), Fibronectin (Thermoscientific; FBN11), CD4 (BioLegend; GK1.5), B220 (BioLegend; RA3–6B2) and cleaved caspase-3 (Cell Signaling; D175), TUNEL staining performed using apoptosis detection kit (Millipore). Images acquired with EVOS FL Auto microscope (Life Technologies). TUNEL+ cells enumerated in 6–10 randomly selected high-powered fields (400X) per slide.
Majumder S., Amatya N., Revu S., Jawale C.V., Wu D., Rittenhouse N., Menk A., Kupul S., Du F., Raphael I., Bhattacharjee A., Siebenlist U., Hand T.W., Delgoffe G.M., Poholek A.C., Gaffen S.L., Biswas P.S, & McGeachy M.J. (2019). IL-17 metabolically reprograms activated fibroblastic reticular cells for proliferation and survival. Nature immunology, 20(5), 534-545.
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