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Ab 100 na

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The AB-100-NA is a piece of lab equipment used for measuring and analyzing samples. It provides accurate and reliable data for research and experimentation purposes.

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Lab products found in correlation

Hk 2 human proximal tubular epithelial cells, by Thermo Fisher Scientific (1 mentions) Tweak, by Merck Group (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Tgf β1, by Thermo Fisher Scientific (1 mentions) Paricalcitol, by Abbott (1 mentions) Ab 406 na, by R&D Systems (1 mentions) Dorsomorphin, by Merck Group (1 mentions) Alk5 inhibitor 2, by Enzo Life Sciences (1 mentions) Sb203580, by Santa Cruz Biotechnology (1 mentions) Sp600125, by Santa Cruz Biotechnology (1 mentions) Nuclear id red green cell viability reagent, by Enzo Life Sciences (1 mentions) Ldn 193189, by Cayman Chemical (1 mentions) Anti rabbit, by Merck Group (1 mentions) Anti mouse, by Merck Group (1 mentions) Mab8929, by R&D Systems (1 mentions) Supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions) Ab 105 c, by R&D Systems (1 mentions) Ab 108 c, by R&D Systems (1 mentions) Rabbit igg, by R&D Systems (1 mentions) Image pro plus, by Media Cybernetics (1 mentions)

8 protocols using ab 100 na

1

Proximal Tubular Epithelial Cell Culture

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HK‐2 human proximal tubular epithelial cells (ATCC, Rockville, MD, USA) were grown on RPMI 1640 (Life Technologies, Grand Island, NY, USA) with 10% heat‐inactivated FBS, 2 mM glutamine, 100 U/ml penicillin, 100 μg/ml streptomycin, 5 μg/ml insulin, 5 μg/ml transferrin, 5 ng/ml sodium selenite and 5 ng/ml hydrocortisone in 5% carbon dioxide at 37°C 17. VHL‐defective clear cell renal carcinoma cells (VHL−/− ccRCC) and VHL+/+ ccRCC cells have been previously described 18. For experiments, cells were rested in serum‐free media 24 hr prior to the addition of stimuli and throughout the experiment. Five hundred thousand cells were seeded in 60 mm diameter wells for RNA extraction, Western blot or flow cytometry experiments. Cells were treated with 100 ng/ml TWEAK (Millipore, Watford, UK), 1 ng/ml TGFβ1 (Peprotech, London, UK), 100 nmol/l paricalcitol (Abbot, Chicago, Illinois), 100 μmol/l TGFβ1 receptor 1 (ALK5) inhibitor (SB431542; Sigma‐Aldrich, Sigma, St. Louis, MO) or 1 ng/ml neutralizing anti‐TGFβ1 antibody (ab100NA; R&D Systems, Minneapolis, MN), based on prior dose–responses studies from our lab.
Poveda J., Sanz A.B., Fernandez‐Fernandez B., Carrasco S., Ruiz‐Ortega M., Cannata‐Ortiz P., Ortiz A, & Sanchez‐Niño M.D. (2016). MXRA5 is a TGF‐β1‐regulated human protein with anti‐inflammatory and anti‐fibrotic properties. Journal of Cellular and Molecular Medicine, 21(1), 154-164.
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2

Neutralizing TGF-β and IL-6 in Mice

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To neutralize TGF-β, mice were treated with a single dose of a polyclonal TGF-β-neutralizing antibody (AB-100-NA; 100 μg/mouse in 100 μl saline; R & D Systems) at 2 d.p.i. by intraperitoneal (i.p.) injection (7 (link)). To neutralize IL-6, mice were treated i.p. with a single dose of a polyclonal antibody to IL-6 (AB-406-NA; 0.5 μg/mouse in 100 μl saline; R & D Systems) at 4 d.p.i. (56 (link)). Controls were treated i.p. with rabbit IgG at both time points (100 μg/mouse in 100 μl saline; Southern Biotech, Birmingham, AL).
Woods P.S., Tazi M.F., Chesarino N.M., Amer A.O, & Davis I.C. (2015). TGF-β-induced IL-6 prevents development of acute lung injury in influenza A virus-infected F508del CFTR-heterozygous mice. American Journal of Physiology - Lung Cellular and Molecular Physiology, 308(11), L1136-L1144.
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3

Pharmacologic Inhibition of TGF-β and MAPK Signaling

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Pharmacologic inhibitors for the TGF-βRI kinase were SB431542 (Calbiochem, Merck, Billerica, MA), activin receptor-like kinase-5 ALK5 Inhibitor I (LY-364947; Enzo Life Sciences AG, Lausen, Switzerland), and ALK5 Inhibitor II (2-(3-(6-Methylpyridin-2-yl)-1H-pyrazol-4-yl)-1,5-naphthyridine; Enzo), all at 10 µM. Pharmacologic inhibitors for the three main MAPK signaling pathways were U0126, SB203580, and SP600125, all at 10 µM (Santa Cruz Biotechnology, SCBT; Santa Cruz, CA). The smad3 inhibitor SIS3 was obtained from Calbiochem. The TGF-β pan specific polyclonal Ab, AB-100-NA was obtained from R&D Systems Inc. (Minneapolis, MN). The bone morphogenetic protein (BMP) type I receptor kinase inhibitors, dorsomorphin (Sigma, St. Louis, MO) and LDN193189 (Cayman, Ann Arbor, MI) were used at 10 µM and 1 µM, respectively. The impact of the TGF-βRI kinase inhibitors on cell viability was determined by the conversion of MTT into formazan crystals [29] (link) and Nuclear-ID Red/Green cell viability reagent (Enzo Life Sciences, Inc., Farmingdale, NY).
Stähli A., Bosshardt D., Sculean A, & Gruber R. (2014). Emdogain-Regulated Gene Expression in Palatal Fibroblasts Requires TGF-βRI Kinase Signaling. PLoS ONE, 9(9), e105672.
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4

Quantifying TGF-β Signaling Inhibition

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Transforming growth factor‐β pan‐specific neutralizing antibody was derived from recombinant human TGFβ1, porcine platelet‐derived TGFβ1.2, porcine platelet‐derived TGFβ2 and recombinant amphibian TGFβ5 porcine platelet‐derived TGFβ1 and 2, which detects TGFβ1,TGFβ1.2, TGFβ2, TGFβ3 and TGFβ5 (AB‐100‐NA, R&D systems). Vascular smooth muscle cells was seeded at 5 × 104 cells/mL in 24 well plates and cultured overnight. The medium was then changed and cells were incubated in the presence or absence of 12 μg/mL TGFβ‐neutralizing antibody for 24 hour. Vascular smooth muscle cells or ECs were fixed and stained with Ki67 antibody as described earlier.
Panahi M., Yousefi Mesri N., Samuelsson E., Coupland K.G., Forsell C., Graff C., Tikka S., Winblad B., Viitanen M., Karlström H., Sundström E, & Behbahani H. (2018). Differences in proliferation rate between CADASIL and control vascular smooth muscle cells are related to increased TGFβ expression. Journal of Cellular and Molecular Medicine, 22(6), 3016-3024.
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5

Western Blot Analysis of RUNX3, TGF-β, and β-actin

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Equal amounts of protein samples (20 μg) were loaded in an 8–12% SDS–polyacrylamide gel and then were transferred to a polyvinyl-iodine fluoride membrane (Biorad, Hercules, CA, USA). After blocking (5% non-fat dry milk), membranes were incubated with primary antibodies overnight at 4 °C (rabbit anti-RUNX3, Abcam ab11905, 1:3000, rabbit anti-TGF-β, R&D systems AB-100-NA, 1:1000, mouse anti-β-actin, R&D systems MAB8929, 1:1000) followed by 1 h incubation with corresponding secondary anti-mouse (1:5000, Sigma) or anti-rabbit (1:5000, Sigma) antibodies conjugated to HRP. Chemiluminescent visualization was performed by SuperSignal West Pico Chemiluminescent Substrate (Thermo Scientific, Waltham, MA, USA) with β-actin as the internal loading control.
Zheng J., Mei Y., Xiang P., Zhai G., Zhao N., Xu C., Liu M., Pan Z., Tang K, & Jia D. (2018). DNA methylation affects metastasis of renal cancer and is associated with TGF-β/RUNX3 inhibition. Cancer Cell International, 18, 56.
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6

Antibody Therapy for Tumor Control

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An antigen affinity-purified polyclonal IgG goat anti-mouse Nrp-1 antibody (AF566) and its isotype control (polyclonal goat IgG) (R&D Systems AB-108-C) were injected i.v. (1.8 mg/kg/mouse) into B16/F10 tumor-bearing mice18 (link). Similar groups of mice were treated i.v. with either a rabbit anti-mouse TGF-β pan-specific polyclonal rabbit antibody (3 mg/kg; R&D Systems AB-100-NA) or an isotype-specific polyclonal rabbit IgG (3 mg/kg; R&D Systems AB-105-C)54 (link). All antibody preparations were reconstituted in sterile PBS and then administered on days +9 and +12 post-B16/F10 implantation. Tumor sizes were monitored every three days (N = 3–6 mice per treatment group depending on the individual experiments presented in the results). The tumor, spleen and TDLN were harvested on day +15 to conduct the in vitro experiments outlined in the results.
Wang S., Gao X., Shen G., Wang W., Li J., Zhao J., Wei Y.Q, & Edwards C.K. (2016). Interleukin-10 deficiency impairs regulatory T cell-derived neuropilin-1 functions and promotes Th1 and Th17 immunity. Scientific Reports, 6, 24249.
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7

Inhibition of TGF-β in Bmi1-deficient Mice

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DCM mice (Bmi1flαMHC-Cretg/+) received intraperitoneal injections of Tgf-β antibody (10 mg per kg body weight in PBS; AB-100-NA; R&D Systems) every 3 days for 2 weeks before being killed. Control mice were treated with identical doses of rabbit IgG (R&D Systems).
Gonzalez-Valdes I., Hidalgo I., Bujarrabal A., Lara-Pezzi E., Padron-Barthe L., Garcia-Pavia P., Gómez-del Arco P., Redondo J.M., Ruiz-Cabello J.M., Jimenez-Borreguero L.J., Enriquez J.A., de la Pompa J.L., Hidalgo A, & Gonzalez S. (2015). Bmi1 limits dilated cardiomyopathy and heart failure by inhibiting cardiac senescence. Nature Communications, 6, 6473.
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8

Intestinal Radiation Injury Evaluation

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Computer-assisted image analysis (Image-Pro Plus, Media Cybernetics, Silver Spring, MD)for histochemistry and immunohistochemistry was used to assess the following established indicators of intestinal radiation injury: 1) proliferation rate of intestinal smooth muscle cells; 2) deposition of collagen in the intestinal wall; and 3) expression of extracellular matrix-associated transforming growth factor-β(TGF-β) as described in detail previously.
Immunohistochemical staining was performed with a standard avidin-biotin complex (ABC) technique, diaminobenzidine (DAB) chromogen, and hematoxylin counterstaining. Appropriate positive (e.g., psoriatic skin for TGF-β) and negative controls (omission of primary antibody and substation of primary antibody with non-immune rabbit IgG) were included. The primary antibodies, incubation times, dilutions, and sources were as follows: monoclonal antibody against proliferating cell nuclear antigen (PCNA) (NA03, 2 hrs, 1:100, Calbiochem, Cambridge, MA) and polyclonal antibody against TGF-β (AB-100-NA, 2 hrs, 1:300 dilution, R&D, Minneapolis, MN).
Wang J., Zheng J., Kulkarni A., Wang W., Garg S., Prather P.L, & Hauer-Jensen M. (2014). Palmitoylethanolamide regulates development of intestinal radiation injury in a mast cell dependent manner. Digestive diseases and sciences, 59(11), 2693-2703.
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