Ab9015

Manufactured by Abcam

Sourced in United Kingdom

Ab9015 is a lab equipment product manufactured by Abcam. It is designed for use in various research applications. The core function of this product is to provide a reliable and consistent tool for researchers to conduct their experiments.

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Lab products found in correlation

5 protocols using ab9015

Quantitative Protein Analysis of Biological Samples

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ConA agarose, HPLC grade acetonitrile (ACN) and formic acid, trifluoroacetic acid, ammonium bicarbonate, iodoacetamide (IAA), and dithiothreitol (DTT) were purchased from Sigma (St. Louis, MO, USA). Sequencing grade trypsin was purchased from Promega (Madison, WI, USA). The 4-plex iTRAQ regents were purchased from ABsciex (Framingham, MA, USA). A TripleTOF 5600 mass spectrometer from ABsciex and an HPLC system from Waters (Milford, MA, USA) were used.
For western blot, the primary antibodies for Alpha-1-antitrypsin (SERRINA1) (SERRINA1, ab9400), Ceruloplasmin (CP, ab51083), Transthyretin (TTR, ab9015), Apolipoprotein A-IV (APOA4, ab81616), Pro-epidermal growth factor (EGF, ab9695) and Prolactin-inducible protein (GCDFP15, ab62363) were purchased from Abcam (Cambridge, UK).

Guo Z., Liu X., Li M., Shao C., Tao J., Sun W, & Li M. (2015). Differential urinary glycoproteome analysis of type 2 diabetic nephropathy using 2D-LC–MS/MS and iTRAQ quantification. Journal of Translational Medicine, 13, 371.

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Immunohistochemical Characterization of PTPRζ

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Most chemicals were purchased from Sigma or Wako Chemicals. The following antibodies were purchased: mouse IgM anti-PTPRζ (sc-33664; Santa Cruz Biotechnology), hereinafter referred to as “anti-PTPRZ (Santa Cruz),” mouse IgM anti-chondroitin sulfate proteoglycan (MAB1581; Merck Millipore), referred to by its clone name, as “Cat-315,” mouse monoclonal anti-aggrecan (6-B-4; Abcam), rabbit polyclonal anti-PTPRZ1(HPA015103; Sigma), hereinafter referred to as “anti-PTPRZ1 (Sigma),” anti-pan cadherin (ab16505; Abcam), polyclonal sheep anti-transthyretin (ab9015; Abcam), referred to as “TTR,” horseradish peroxidase (HRP)-conjugated goat anti-mouse IgM (SAB-110; Stressgen), goat anti-rabbit IgG (NA934; GE Healthcare), and donkey anti-sheep IgG-HRP (A16041; Thermo Fisher Scientific).

Yamanoi Y., Fujii M., Murakami Y., Nagai K., Hoshi K., Hashimoto Y., Honda T., Saito K, & Kitazume S. (2020). Soluble protein tyrosine phosphatase receptor type Z (PTPRZ) in cerebrospinal fluid is a potential diagnostic marker for glioma. Neuro-oncology Advances, 2(1), vdaa055.

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Western Blotting Protocol for Protein Analysis

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Western blotting was conducted as previously described [4 (link)]. Samples for mass spectrometry were also used for western blotting after TEAB buffer was added. For this study, a primary monoclonal mouse anti-β-actin antibody 1:5,000 (clone AC-15, Sigma-Aldrich, Søborg, Denmark) and a primary polyclonal sheep anti-transthyretin (TTR) antibody 1:5,000 (ab9015, Abcam, Cambridge, UK) were used. Both antibodies were diluted in 2.5% (w/v) skim milk blocking buffer. Horseradish peroxidase (HRP)–conjugated secondary antibodies polyclonal goat anti-mouse immunoglobulins/HRP (Dako, Glostrup, Denmark) and rabbit anti-sheep IgG eavy and light chains (ab6747, Abcam, Cambridge, UK) were used, diluted 1;30,000 and 1:1;000, respectively, in 2.5% (w/v) skim milk blocking buffer.

Cehofski L.J., Kruse A., Alsing A.N., Nielsen J.E., Pedersen S., Kirkeby S., Honoré B, & Vorum H. (2018). Intravitreal bevacizumab upregulates transthyretin in experimental branch retinal vein occlusion. Molecular Vision, 24, 759-766.

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Western Blot Analysis of Rat Knee Tissues

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Rat knee joint tissues were homogenized and lysed in a protein extraction reagent (Tissue protein extraction reagent, Pierce, Rockford, US). Western blot analysis was performed as described previously [29 (link)]. The primary antibodies to transferrin (17435-1-AP; Proteintech), catalase (ab209211; Abcam), SOD1 (ab16831; Abcam), SOD2 (ab68155; Abcam), SOD3 (ab21974; Abcam), glutathione peroxidase 1 (ab59546; Abcam), glutathione peroxidase 2 (MAB5470; R&D Systems), transthyretin (ab9015; Abcam), NF-κB (8242S; Cell Signaling), IL-1β (ab9722; Abcam), RANKL (sc-9073; Santa Cruz), and GAPDH (sc-20357; Santa Cruz) were obtained commercially. After extensive washing, appropriate secondary antibodies were used in the experiments, including anti-rabbit IgG antibody conjugated with horseradish peroxidase (7074S; Cell Signaling), anti-mouse IgG antibody conjugated with horseradish peroxidase (7076P2; Cell Signaling), or anti-sheep IgG antibody conjugated with horseradish peroxidase (313-035-003; Jackson ImmunoResearch).

Peng K.T., Tsai M.H., Lee C.W., Chiang Y.C., Chen P.C., Chen C.C., Chang C.H., Shih H.N, & Chang P.J. (2018). Dysregulated expression of antioxidant enzymes in polyethylene particle-induced periprosthetic inflammation and osteolysis. PLoS ONE, 13(8), e0202501.

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Immunostaining of Retinal BRVO Samples

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Eyes from one animal were used for immunohistochemistry to compare BRVO + bevacizumab (n=1) to BRVO + NaCl (n=1). Immunohistochemical analyses were performed on retinal sections containing the occlusions. Fixation was performed using a fixative of 99% ethanol (three parts) and glacial acetic acid (one part) as previously described [4 (link)]. A polyclonal IgG antibody directed at TTR (ab9015, Abcam) was used for immunohistochemistry. Sheep antibodies were diluted (1:200 to 1:800) in PBS (1X; 137 mM NaCl, 2.7 mM KCl, 10 mM Na₂HPO₄, 1.8 mM KH₂PO₄, pH 7.4) with 0.3% Triton X100. Retinal sections were incubated overnight at 4 °C and were processed using EnVission (DakoCytomation, Glostrup, Denmark) DAB. Controls were incubated with either sheep IgG_2b alone or irrelevant sheep antibodies.

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