Ph d 12 phage display library

Manufactured by New England Biolabs

Sourced in China, United States

The Ph.D.-12 phage display library is a tool for the screening and selection of peptide sequences. It consists of a diverse library of 12-mer peptides displayed on the surface of M13 bacteriophage particles. This library can be used to identify peptides that bind to a target of interest through an affinity-based selection process.

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Lab products found in correlation

Dynabeads protein g, by Thermo Fisher Scientific (1 mentions) Anti flag m2 monoclonal antibody, by Merck Group (1 mentions) Dynabead, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Ph.D.-12TM Phage Display Peptide Library Ph.D-12TM Phage Display Peptide Library Kit Ph.D.-12

4 protocols using ph d 12 phage display library

Phage Display Library Screening for Antibody Binding Peptides

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The Ph.D-12 phage display library was purchased from New England Biolabs. Subsequently, this library was amplified on approximately 200 solid plates, each 150 mm in diameter. According to the protocol, the amplified Ph.D-12 phage display library was purified, titered, and stored at −20 °C in 50% glycerin. For screening of antibodies’ binding peptides, each well of a 96-well PCR plate was blocked with 200 μl of 3% bovine serum albumin in TBST (containing 0.5% Tween 20 in TBS) overnight on a rotator at 4 °C. Immunoglobulin G (IgG) antibodies (0.02–0.8 μg) of and 10 μl of the amplified PhD.-12 library (∼100 fold representation, 10¹¹ plaque-forming units for a library of 10⁹ clones) were added to each preblocked well. The antibodies and phages from the PhD.-12 library were incubated overnight on a rotator at 4 °C. Ten microliters of Dynabeads protein G (Thermo Scientific) was added into each well and incubated for 4 h on a rotator at 4 °C to capture the antibody–phage complex. With a 96-well magnetic stand, the Dynabeads protein G was washed three times with 200 μl of TBST, followed by an additional wash with deionized water. The beads in each well were resuspended in 15 μl of deionized water, and the phages were lysed at 95 °C for 10 min. The phage lysates were stored at −80 °C.

Qi H., Ma M., Hu C., Xu Z.W., Wu F.L., Wang N., Lai D.Y., Li Y., Zhang H., Jiang H.W., Meng Q.F., Guo S., Kang Y., Zhao X., Li H, & Tao S.C. (2021). Antibody Binding Epitope Mapping (AbMap) of Hundred Antibodies in a Single Run. Molecular & Cellular Proteomics : MCP, 20, 100059.

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Phage Display Screening of hBMSCs

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Peptide sequences with affinity for hBMSCs were identified by screening the Ph.D.12^™ Phage Display Library (New England Biolabs, #E8110S), consisting of 10⁹ different phage with 12-mer amino acid linear peptide inserts, against clonally derived hBMSCs (Passage 3–6). 2×10⁴ hBMSCs were plated in 2 wells of a 6-well dish and cultured for 6 days in culture media at 37°C and 5% CO₂. After 6 days of culture, the NEB 12-mer peptide library was prescreened against sintered HAP disks prior to introduction to the cells, to preferentially screen for sequences attracted to the cells and not the apatite. A streptavidin control was run in a separate dish per the manufacturer’s protocol.
Plated cells in 6-well dishes were rinsed 2X with phosphate buffered saline (PBS, Gibco #10010) and pre-blocked with α-MEM containing 0.1% bovine serum albumin (BSA) without supplements at 37°C and 5% CO₂ for 30 minutes. The aliquot of phage harvested from the HAP disks was then introduced to the cells (n=2). Non-binding phage were then discarded, and the cells were washed 5X in cold PBS. The phage bound to the cells was eluted with 1 mL of Glycine/HCl, pH 2.2, with 1 mg/mL BSA for 10 minutes at room temperature while being gently rocked. The eluted phage was collected and neutralized with 1M Tris-HCl, pH 9.1. Three rounds of panning were performed for each sample.

Ramaraju H., Miller S.J, & Kohn D.H. (2017). Dual-functioning peptides discovered by phage display increase the magnitude and specificity of BMSC attachment to mineralized biomaterials. Biomaterials, 134, 1-12.

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Phage Display Screening for DC-Targeting Peptides

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The Ph.D.-12 phage display library (NEB, Beijing, China) displaying linear 12-mer random peptide at the N-terminus of P III protein of bacteriophage M 13 was applied to screen the DC-targeting peptide. First round: phages (2 × 10¹¹) were added into chBM-DCs for 30 min at 4°C. The cell suspensions centrifuged at 600 × g for 8 min were resuspended in PBS containing 1% bovine serum albumin (BSA) and 0.05% Tween-20. After repeating the washing step three times, the number of phages bound to DCs was evaluated by the phage-plaque assay. The phages were amplified in Escherichia coli ER 2,738 for the next round of biopanning. Second-four rounds: phages were incubated with marrow cells and unbound phages were added into chBM-DCs for 15 min at 4°C, and then the procedures described above were repeated. After the fourth round of biopanning, individual phage-plaques were randomly selected and amplified separately. The nucleotide sequence of each phage extracted with the Phage DNA Isolation Kit (EasyExtraction, Beijing, China) was determined with −96 g III primer, and the sequence was translated into a peptide sequence.

Ma S., Qiao X., Xu Y., Wang L., Zhou H., Jiang Y., Cui W., Huang X., Wang X., Tang L, & Li Y. (2019). Screening and Identification of a Chicken Dendritic Cell Binding Peptide by Using a Phage Display Library. Frontiers in Immunology, 10, 1853.

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Phage Display Selection of PD-L1 Binders

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We performed two rounds of phage display selection using recombinant human PD-L1 extracellular domain (ECD) protein (Cat# 10084-H08H, Sino Biological Inc., Beijing, China) as bait. The selection of Ph.D.-12 phage display library (New England Biolabs, Ipswitch, MA, United States) against PD-L1 was performed in six replicates. The control selections, i.e., Ph.D.-12 against Dynabeads (Cat# 10-103-D, Invitrogen) and Ph.D.-12 against unrelated anti-FLAG M2 monoclonal antibody (Cat# F3165, Sigma-Aldrich), were performed in triplicate.

He B., Li B., Chen X., Zhang Q., Lu C., Yang S., Long J., Ning L., Chen H, & Huang J. (2022). PDL1Binder: Identifying programmed cell death ligand 1 binding peptides by incorporating next-generation phage display data and different peptide descriptors. Frontiers in Microbiology, 13, 928774.

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