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Pepstar peptide microarrays

Manufactured by JPT Peptide Technologies
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PepStar peptide microarrays are a tool for high-throughput screening and analysis of peptide-based interactions. The core function of the PepStar system is to enable the immobilization of peptides on a solid support, allowing for the simultaneous detection and quantification of multiple peptide-protein or peptide-peptide interactions.

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Lab products found in correlation

Leupeptin, by Omni International (2 mentions) Aprotinin, by Omni International (2 mentions) Phenylmethylsulphonyl fluoride, by Omni International (2 mentions) Bead ruptor homogenizer, by Omni International (2 mentions) Powerscanner microarray scanner, by Tecan (2 mentions) Tissueruptor, by Qiagen (1 mentions)

7 protocols using pepstar peptide microarrays

1

Epitope Mapping of Murine ApoA-I

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Serum samples from the mice shown in Figs. 2 and 3 with an anti–full-length ApoA-I absorbance (A450) >1.0 at study termination were selected for epitope mapping by peptide array. Where possible, a matched sample from 11 wk of age was also included. Serum samples were analyzed by JPT Peptide Technologies (Berlin, Germany) PepStar peptide microarrays using a series of 58 synthetic 15-mer peptides with 4-aa offsets spanning the entire 243-aa sequence of murine ApoA-I. Serum samples were incubated with the microarray slide for 1 h at 30°C. After washing the microarray, fluorescently labeled anti-mouse IgG secondary Ab at 1 μg/ml was added and incubated for 1 h. Control wells lacking mouse serum were used to assess nonspecific binding of the secondary Ab toward each peptide. After washing, the array was scanned at 635 nm to obtain fluorescence intensity profiles and resulting images were quantified to yield a mean pixel value for each peptide, which is represented as a heatmap.
Pitts M.G., Nardo D., Isom C.M, & Venditto V.J. (2020). Autoantibody Responses to Apolipoprotein A-I Are Not Diet- or Sex-Linked in C57BL/6 Mice. ImmunoHorizons, 4(8), 455-463.
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2

Peptide Array Protocol for Jejunum Tissue

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PepStar peptide microarrays were obtained from JPT Peptide Technologies GmbH (Berlin, Germany), and the peptide array protocol was carried out as previously described (6 (link)) with the following modifications (8 (link), 17 (link)). Jejunum tissue samples were weighed to obtain a consistent 40 mg sample for the array protocol. Samples were homogenized by a hand-held TissueRuptor (Qiagen, Valencia, CA, USA) in 100 µL of lysis buffer (20 mM Tris–HCl pH 7.5, 150 mM NaCl, 1 mM Ethylenediaminetetraacetic acid, 1 mM ethylene glycol tetraacetic acid, 1% Triton X-100, 2.5 mM sodium pyrophosphate, 1 mM Na3VO4, 1 mM NaF, 1 µg/mL leupeptin, 1 g/mL aprotinin, and 1 mM Phenylmethylsulphonyl fluoride). All chemicals purchased from Sigma-Aldrich, Co. (St. Louis, MO, USA) unless otherwise indicated.
Swaggerty C.L., Kogut M.H., He H., Genovese K.J., Johnson C, & Arsenault R.J. (2017). Differential Levels of Cecal Colonization by Salmonella Enteritidis in Chickens Triggers Distinct Immune Kinome Profiles. Frontiers in Veterinary Science, 4, 214.
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3

HER2 Antibody Response Profiling

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Serum collections were analyzed for HER2 peptide-specific antibody response using a panel of overlapping 15-mer peptides spanning the HER2 sequence using PepStar peptide microarrays, Multiwell Microarray Service (JPT Peptide Technologies GmbH, Germany). The reactivity pattern was summarized in fold increase compared with baseline. As this is a study conducted in non-HLA preselected patients with a small number of available biospecimens, responses to multiple epitopes in individual patients or detection of varying epitopes among patients will be reported in a descriptive manner.
Maeng H.M., Moore B.N., Bagheri H., Steinberg S.M., Inglefield J., Dunham K., Wei W.Z., Morris J.C., Terabe M., England L.C., Roberson B., Rosing D., Sachdev V., Pack S.D., Miettinen M.M., Barr F.G., Weiner L.M., Panch S., Stroncek D.F., Wood L.V, & Berzofsky J.A. (2021). Phase I Clinical Trial of an Autologous Dendritic Cell Vaccine Against HER2 Shows Safety and Preliminary Clinical Efficacy. Frontiers in Oncology, 11, 789078.
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4

Peptide Array Profiling of Gut Samples

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Peptide array protocol was carried out as previously [14 (link)] described and summarized below using PepStar peptide microarrays from JPT Peptide Technologies GmbH (Berlin, Germany). Ileum and jejunum tissue samples were weighed and a 40 mg sample was homogenized by a Bead Ruptor homogenizer (Omni, Kennesaw GA) in 100 μL of lysis buffer (20 mM Tris–HCl pH 7.5, 150 mM NaCl, 1 mM Ethylenediaminetetraacetic acid (EDTA), 1 mM ethylene glycol tetraacetic acid (EGTA), 1% Triton X-100, 2.5 mM sodium pyrophosphate, 1 mM Na3VO4, 1 mM NaF, 1 μg/mL leupeptin, 1 g/mL aprotinin and 1 mM Phenylmethylsulphonyl fluoride). All chemicals were purchased from Sigma-Aldrich, Co. (St. Louis, MO) unless specified otherwise. Arrays were then imaged using a Tecan PowerScanner microarray scanner (Tecan Systems, San Jose, CA, USA) at 532–560 nm with a 580 nm filter to detect dye fluorescence.
Swaggerty C.L., Arsenault R.J., Johnson C., Piva A, & Grilli E. (2020). Dietary supplementation with a microencapsulated blend of organic acids and botanicals alters the kinome in the ileum and jejunum of Gallus gallus. PLoS ONE, 15(7), e0236950.
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5

Peptide Array Analysis of Jejunum Proteins

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Peptide array protocol was carried out as previously described (Arsenault et al., 2017 (link)) and summarized below using PepStar peptide microarrays from JPT Peptide Technologies GmbH (Berlin, Germany). A 40 mg sample of jejunum was homogenized by a Bead Ruptor homogenizer (Omni, Kennesaw, GA) in 100 μL of lysis buffer (20 mM Tris–HCl pH 7.5, 150 mM NaCl, 1 mM Ethylenediaminetetraacetic acid (EDTA), 1 mM ethylene glycol tetraacetic acid (EGTA), 1% Triton X-100, 2.5 mM sodium pyrophosphate, 1 mM Na3VO4, 1 mM NaF, 1 μg/mL leupeptin, 1 g/mL aprotinin and 1 mM Phenylmethylsulphonyl fluoride). All chemicals were purchased from Sigma-Aldrich, Co. (St. Louis, MO) unless specified otherwise. Arrays were then imaged using a Tecan PowerScanner microarray scanner (Tecan Systems, San Jose, CA) at 532 to 560 nm with a 580 nm filter to detect dye fluorescence.
Swaggerty C.L., Byrd JA I.I., Arsenault R.J., Perry F., Johnson C.N., Genovese K.J., He H., Kogut M.H., Piva A, & Grilli E. (2022). A blend of microencapsulated organic acids and botanicals reduces necrotic enteritis via specific signaling pathways in broilers. Poultry Science, 101(4), 101753.
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6

Epitope Mapping of Murine ApoA-I

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Serum samples from the mice shown in Figs. 2 and 3 with an anti–full-length ApoA-I absorbance (A450) >1.0 at study termination were selected for epitope mapping by peptide array. Where possible, a matched sample from 11 wk of age was also included. Serum samples were analyzed by JPT Peptide Technologies (Berlin, Germany) PepStar peptide microarrays using a series of 58 synthetic 15-mer peptides with 4-aa offsets spanning the entire 243-aa sequence of murine ApoA-I. Serum samples were incubated with the microarray slide for 1 h at 30°C. After washing the microarray, fluorescently labeled anti-mouse IgG secondary Ab at 1 μg/ml was added and incubated for 1 h. Control wells lacking mouse serum were used to assess nonspecific binding of the secondary Ab toward each peptide. After washing, the array was scanned at 635 nm to obtain fluorescence intensity profiles and resulting images were quantified to yield a mean pixel value for each peptide, which is represented as a heatmap.
Pitts M.G., Nardo D., Isom C.M, & Venditto V.J. (2020). Autoantibody Responses to Apolipoprotein A-I Are Not Diet- or Sex-Linked in C57BL/6 Mice. ImmunoHorizons, 4(8), 455-463.
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7

Fibrinogen and Autoantigen Peptide Microarray

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PepStar™ peptide microarrays (JPT Peptide Technologies, Berlin, Germany) comprise 169 15-mers from the alpha and beta chain of fibrinogen, vimentin, histon 4, enolase, proteoglycan, filaggrin, ebna 2 and collagen (Supplementary Figures 1, 2). Thirty three arginine-containing peptides and their 136 citrulline-substituted variants (Supplementary Table 1) were synthesized on cellulose membranes using SPOT™ synthesis technology (9 (link)). A reactivity tag was attached to the N-terminus of each peptide. The peptides were cleaved and eluted from the membrane. Quality control measurements using liquid chromatography mass spectrometry were performed. Peptides were immobilized on the epoxy-modified slide surfaces using chemoselective coupling. All peptides were deposited (10−15 moles) in three identical subarrays per block/slide enabling analysis of assay homogeneity and reliability of the results. Peptide microarrays were scanned after the printing process for identification and quality control of each individual spot.
Hemon M.F., Lambert N.C., Arnoux F., Roudier J, & Auger I. (2022). PAD4 Immunization Triggers Anti-Citrullinated Peptide Antibodies in Normal Mice: Analysis With Peptide Arrays. Frontiers in Immunology, 13, 840035.
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