Tentagel r ram resin

Manufactured by Rapp Polymere

Sourced in Germany

TentaGel R RAM resin is a solid-phase synthesis resin used in laboratory applications. It is a polyethylene glycol-grafted polystyrene resin with a Rink amide linker. The resin supports the immobilization of chemical compounds for various synthesis and analysis purposes.

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Lab products found in correlation

5 6 carboxyfluorescein, by Merck Group (2 mentions) Piperidine, by Merck Group (2 mentions) Prelude x, by Protein Technologies (1 mentions) Api 2000, by Thermo Fisher Scientific (1 mentions) Ultimate 3000, by Phenomenex (1 mentions) Triisopropylsilane, by Merck Group (1 mentions) Hydrazine monohydrate, by Merck Group (1 mentions) Acetonitrile acn, by Avantor (1 mentions) Dichloromethane dcm, by Biosolve (1 mentions) Nα fmoc protected amino acids, by Iris Biotech (1 mentions) N n diisopropylcarbodiimide, by Iris Biotech (1 mentions) 4 methylmorpholine, by Merck Group (1 mentions) Diethyl ether, by Merck Group (1 mentions) Atto488, by ATTO-TEC (1 mentions) Dimethylformamide dmf, by Biosolve (1 mentions) Fmoc protected amino acids, by Merck Group (1 mentions) Anhydrous hydrazine, by Merck Group (1 mentions) N n diisopropylcarbodiimide, by Merck Group (1 mentions) Fmoc l cys trt oh, by Gyros Protein Technologies (1 mentions) Fmoc l lys boc oh, by Gyros Protein Technologies (1 mentions)

9 protocols using tentagel r ram resin

Synthetic Peptide Sequence Preparation

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((GPO)₅(G*P*O)(GPO)₅)₃ was synthesized as previously reported⁷. Briefly, the peptide was synthesized (0.1 mmol scale) as C-terminal amides on a TentaGel R RAM resin (loading of 0.19 mmol/g, Rapp Polymere) following the standard Fmoc-based solid-phase peptide synthesis strategy on a microwave-assisted automated peptide synthesizer (Liberty™, CEM), and purified by reverse phase HPLC. Pure peptides were characterised by matrix-assisted laser desorption and ionization-time of flight (MALDI-TOF) mass spectrometry (Supplementary Fig. S1).

Chow W.Y., Forman C.J., Bihan D., Puszkarska A.M., Rajan R., Reid D.G., Slatter D.A., Colwell L.J., Wales D.J., Farndale R.W, & Duer M.J. (2018). Proline provides site-specific flexibility for in vivo collagen. Scientific Reports, 8, 13809.

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Solid-Phase Peptide Synthesis Protocol

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Peptides were synthesized on a Prelude X instrument (Protein Technologies, Inc., Tucson, AZ) using standard Fmoc solid-phase peptide synthesis at 25 μmole scale. Synthesis-grade Fmoc amino acid (AA) reagents were purchased from Protein Technologies, Inc. or AAPPTec (Louisville, KY). Standard coupling conditions comprised addition of 0.65 mL Fmoc-AA (200 mM in DMF), 0.65 mL HATU (195 mM in DMF), and 0.5 mL NMM (0.6 M in DMF). Coupling reactions were mixed for 25 min at 50°C. Fmoc deprotection employed 20% piperidine in DMF. To generate C-terminal carboxamides, peptides were prepared on TentaGel R RAM resin (Rapp-Polymere, Tübingen, Germany). N-terminal fluorescein capping was performed with 5 (6)-carboxyfluorescein (Sigma-Aldrich, St. Louis, MO) while acetyl capping utilized acetic anhydride with 0.6 M NMM. Peptide resin was thoroughly washed with DCM and dried under vacuum after synthesis was complete. Cleavage of the peptide from resin was performed with 4 mL [92.5/2.5/2.5/2.5%] of TFA/water/TIS/EDT for 3 hr. The TFA solution was then precipitated in ice-cold ether and centrifuged at 5100 RPM. Supernatant was decanted while pellets were washed twice more and then dried overnight under vacuum.

Sdano M.A., Fulcher J.M., Palani S., Chandrasekharan M.B., Parnell T.J., Whitby F.G., Formosa T, & Hill C.P. (2017). A novel SH2 recognition mechanism recruits Spt6 to the doubly phosphorylated RNA polymerase II linker at sites of transcription. eLife, 6, e28723.

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Automated Synthesis of Zinc Finger Peptides

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Zinc fingers (ZFs): ZScan20-2, ZNF442, consensus ZF CP1 (CCHH), ZF133-11, and its C7E mutant (ZF133-11 C7E) with weaker Zn(II) affinity were synthesized on TentaGel R RAM resin (loading 0.19 mmol/g, Rapp Polymere GmbH) using a Liberty 1 microwave-assisted synthesizer (CEM, USA) according to the standard Fmoc strategy described in detail before.^{52 (link),53 (link)} The full sequences are given in Table S1. All peptides were acetylated at the N-terminus and purified by HPLC (Dionex Ultimate 3000 system) on a Phenomenex C18 column (Gemini-NX 5 µm, 110 Å) using a gradient of 95% acetonitrile/5% water (0.1% TFA) with 0.1% TFA in water, as described previously.^{53 (link)} After purification, peptides were lyophilized. The identity of the obtained pure peptides was confirmed by ESI-MS (API 2000, Applied Biosystems, USA). The list of theoretical and experimental masses is presented in Table S2.

Pomorski A., Drozd A., Kocyła A, & Krężel A. (2023). From methodological limitations to the function of metallothioneins - a guide to approaches for determining weak, moderate, and tight affinity zinc sites. Metallomics: Integrated Biometal Science, 15(5), mfad027.

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Microwave-Assisted Peptide Synthesis and Purification

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Peptides were synthesised (0.1 mmol scale) as C-terminal amides on a TentaGel R RAM resin (loading of 0.19 mmol/g, Rapp Polymere) following the standard Fmoc-based solid-phase peptide synthesis strategy on a microwave-assisted automated peptide synthesiser (Liberty^TM, CEM), and purified by reverse phase HPLC as detailed in Supplementary Information, Section S1.
Pure peptides were characterised by matrix-assisted laser desorption and ionization-time of flight (MALDI-TOF) mass spectrometry, then lyophilised.

Ying Chow W., Bihan D., Forman C.J., Slatter D.A., Reid D.G., Wales D.J., Farndale R.W, & Duer M.J. (2015). Hydroxyproline Ring Pucker Causes Frustration of Helix Parameters in the Collagen Triple Helix. Scientific Reports, 5, 12556.

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Solid-Phase Peptide Synthesis Reagents

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N‐α‐Fmoc‐protected amino acids, ethyl 2‐cyano‐2‐(hydroxyimino)acetate (Oxyma), and N,N′‐diisopropylcarbodiimide (DIC) were purchased from Iris Biotech (Marktredwitz, Germany), and F‐Wang resin, NovaSyn® W‐TGA resin, O‐(7‐azabenzotriazolyl)‐tetramethyluronium hexafluorophosphate (HATU), and O‐(1H‐6‐chlorobenzotriazole‐1‐yl)‐1,1,3,3‐tetramethyluronium hexafluorophosphate (HCTU) were supplied from Novabiochem (Darmstadt, Germany). TentaGel R RAM resin was obtained from Rapp Polymere (Tübingen, Germany). 6‐Carboxytetramethylrhodamine (TAMRA) was purchased from ChemPep, Inc. (Wellington, Florida). Atto488 and Atto565 dyes were obtained from ATTO TEC (Siegen, Germany). Acetonitrile (ACN) was purchased from VWR (Darmstadt, Germany). Dimethylformamide (DMF) and dichloromethane (DCM) were obtained from Biosolve (Valkenswaard, The Netherlands). N,N‐Diisopropylethylamine (DIPEA), 1,2‐ethanedithiol (EDT), 4‐(2‐hydroxyethyl)‐1‐piperazineethanesulfonic acid (HEPES), hydrazine monohydrate, 4‐methoxytriphenylmethyl chloride (Mmt‐Cl), mercaptophenyl acetic acid (MPAA), 4‐methylmorpholine (NMM), piperidine, thioanisole (TA), trifluoroacetic acid (TFA), and triisopropylsilane (TIS) were purchased from Sigma‐Aldrich (Taufkirchen, Germany). Diethyl ether was from Merck (Darmstadt, Germany).

Wolf P., Mohr A., Gavins G., Behr V., Mörl K., Seitz O, & Beck‐Sickinger A.G. (2021). Orthogonal Peptide‐Templated Labeling Elucidates Lateral ETAR/ETBR Proximity and Reveals Altered Downstream Signaling. Chembiochem, 23(6), e202100340.

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Solid-Phase Peptide Synthesis and Purification

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The peptides were prepared by solid-phase peptide synthesis using the standard cycles of a Millipore 9050 automatic peptide synthesizer (Millipore, Darmstadt, Germany) and Fmoc chemistry. Fmoc-protected amino acids from NovaBiochem (Merck Millipore, Darmstadt, Germany) were used at four-fold excess, the Tentagel-R-RAM resin was from Rapp Polymere (Tübingen, Germany). Cleavage of the peptide was performed in 28 mL trifluoroacetic acid (TFA), 1.5 mL water and 0.3 mL triethylsilane for 4 h. After solubilization and ether precipitation the TFA counter ions were exchanged in 4% acetic acid. After purification by semipreparative HPLC using an acetonitrile/water gradient on a C18 reverse-phase column (Luna 5u c18(2) 100 Å; 150 × 30 mm, Phenomenex, Le Pecq, France) the identity of the products was confirmed by MALDI TOF mass spectrometry.

Aisenbrey C., Rifi O, & Bechinger B. (2020). Structure, membrane topology and influence of cholesterol of the membrane proximal region: transmembrane helical anchor sequence of gp41 from HIV. Scientific Reports, 10, 22278.

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Solid-Phase Peptide Synthesis with HATU

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2-chlorotrityl resin and 1-[Bis(dimethylamino)methylene]−1 H-1,2,3-triazolo[4,5-b]pyridinium-3-oxidhexafluorophosphate (HATU) were purchased from ChemPep. TentaGel R RAM resin was purchased from Rapp Polymere. Boc-Cys(Trt)-OH, Fmoc-L-Cys(Trt)-OH, Fmoc-L-Lys(Boc)-OH, Fmoc-L-Asp(tBu)-OH, Fmoc-L-Glu(tBu)-OH, Fmoc-L-Ser(tBu)-OH, Fmoc-L-Asn(Trt)-OH, Fmoc-Gly-OH, Fmoc-L-Ala-OH, Fmoc-L-Val-OH, Fmoc-L-Leu-OH, and Fmoc-Gly-Ser(psiMe,Mepro)-OH were purchased from Gyros Protein Technologies. Fmoc-Lys(Ac)-OH was purchased from Anaspec. Fmoc-Lys(Dde)-OH was purchased from AAPPTec. Synthesis grade trifluoracetic acid (TFA), ACS grade dimethylformamide (DMF), peptide synthesis grade n-methylmorpholine (NMM), synthesis grade n-methylpyrrolidinone (NMP), ACS grade anhydrous diethyl ether, HPLC grade acetonitrile (ACN), HPLC grade methanol, LC-MS grade ACN with 0.1% formic acid, and LC-MS grade water with 0.1% formic acid were purchased from Fisher Scientific (all reagent brands from Fisher Scientific were Fisher Chemical). Piperidine, triisopropylsilane (TIS), 1,2-ethanedithiol (EDT), 5 (6)-carboxyfluorescein, N,N′-Diisopropylcarbodiimide, Oxyma Pure, anhydrous hydrazine, acetic anhydride, and 4-mercaptophenylacetic acid (MPAA) were purchased from Sigma Aldrich.

Han H., Fulcher J.M., Dandey V.P., Iwasa J.H., Sundquist W.I., Kay M.S., Shen P.S, & Hill C.P. (2019). Structure of Vps4 with circular peptides and implications for translocation of two polypeptide chains by AAA+ ATPases. eLife, 8, e44071.

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Preparation of Chiral Precursors and Compound Library Synthesis

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Deuterated (S)-(−)-2-bromopropionic acid, N-Alloc-1,4-ethylenediamine hydrochlroride, and (S)-2-chloro-3-phenylpropanoic acid were prepared as described previously.^{22 (link)} All the other chemicals were purchased from commercial sources and used without further purification. (S)-(−)-Chloropropionic acid was purchased from Alfa Aeser. (S)-2-Chloro-4-methylvaleric acid was purchased from TCI America. TentaGel R RAM resin was purchased from Rapp Polymere. All the Fmoc-protected amino acids and Knorr Amide MBHA resin were purchased from Novabiochem. All the other chemicals were obtained from Sigma-Aldrich, Acros Organics, or Oakwood Chemical. Commercial sources of materials required for screening are described in the screening section. All MALDI-TOF and TOF/TOF mass spectra were obtained using 4800 Plus MALDI TOF/TOF Analyzer (Applied Biosystems). Cyano-4-hydroxycinnamic acid was used as a matrix for the mass analysis. Phosphate-buffered saline (PBS) solutions were diluted from a 10× stock PBS solution (Corning). Tris-buffered saline (TBS) solutions were diluted from a 20× TBS stock (Thermo Scientific). The following section details experimental procedures for serum preparation and autoantibody binding measurements. A descriptive experimental flow for library synthesis is provided in the supplementary information.

Doran T.M., Morimoto J., Simanski S., McEnaneya P.J, & Kodadek T. (2015). Reliable Diagnosis of Murine Type 1 Diabetes Using A Panel of Autoantigens and “Antigen Surrogates” Mounted Onto A Liquid Array. Molecular bioSystems, 11(11), 3156-3163.

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Fmoc-Amino Acid Peptide Synthesis

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Reagents All Fmoc-amino acids were obtained from GL Biochem (Shanghai, China), or Matrix Innovation (Quebec City, Canada), with the following side-chain protecting groups: Asn (Trt), His(Trt), Ser(tBu), Thr(tBu), Tyr(tBu). Rink Amide MBHA resin (0.48 mmol g -1 ) was purchased from Matrix Innovation (Quebec City, Canada). TentaGel R RAM resin (0.18 mmol g -1 ) was purchased from Rapp Polymere GmbH (Tübingen, Germany). This article is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported Licence.

Strauss P., Nuti F., Quagliata M., Papini A.M, & Hurevich M. (2023). Accelerated solid-phase synthesis of glycopeptides containing multiple N-glycosylated sites. Organic & biomolecular chemistry, 21(8).

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