Log In Sign up
The largest database of trusted experimental protocols

Model 433a peptide synthesizer

Manufactured by Thermo Fisher Scientific
Visit Supplier
Sourced in United States

The Model 433A peptide synthesizer is a laboratory instrument designed for the automated synthesis of peptides. It provides a controlled and efficient process for the step-by-step assembly of peptide chains from individual amino acid building blocks.

Automatically generated - may contain errors

Lab products found in correlation

Pierce fab preparation kit, by Thermo Fisher Scientific (1 mentions) Beckman du 640 spectrophotometer, by Beckman Coulter (1 mentions) Sodium hydroxide (naoh), by Thermo Fisher Scientific (1 mentions) 1 cm quartz cuvette, by Hellma (1 mentions) Monos ion exchange column, by GE Healthcare (1 mentions) Hitrap protein g column, by GE Healthcare (1 mentions) Fab preparation kit, by Thermo Fisher Scientific (1 mentions)

6 protocols using model 433a peptide synthesizer

1

Synthetic Peptide Preparation and Fab Generation

Cited 3 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
All peptides were chemically synthesized by the Core Laboratory of the Center for Biologics Evaluation and Research at the US Food and Drug Administration, by using an Applied Biosystems model 433A peptide synthesizer as previously described (22 (link), 33 (link)). Ascites containing mAb1H8 was produced by Harlan Bioproducts for Science as previously described (18 (link)). Fab of the immunoglobulin molecule was prepared using the Pierce Fab Preparation Kit (44985) from Thermo Fisher Scientific by following the instructions from the manufacturer.
Deng L., Hernandez N., Zhong L., Holcomb D.D., Yan H., Virata M.L., Tarafdar S., Xu Y., He Y., Struble E., Alter H.J, & Zhang P. (2021). A conserved epitope III on hepatitis C virus E2 protein has alternate conformations facilitating cell binding or virus neutralization. Proceedings of the National Academy of Sciences of the United States of America, 118(28), e2104242118.
+ Open protocol
+ Expand
2

Peptide Synthesis and Characterization

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Peptide synthesis was conducted on an Applied Biosystems model 433A peptide synthesizer as previously described (Lee et al., 2010 (link)). The linear precursor was synthesized using solid-phase methodology with Fmoc chemistry, starting from Fmoc-Pro-Wang resin (Fmoc-Thr-Wang resin for PcTx1) using a variety of blocking groups for the protection of the amino acids. All the GxTx1E peptides were synthesized with norleucine (Nle) in place of Met35 to avoid oxidation of the toxin. A 4 mol excess of Fmoc amino acid, DIC, and Cl-HOBt were used for amino acid activation. After trifluoroacetic acid cleavage, the crude linear peptide was extracted with 2 M acetic acid and diluted to a final concentration of 25 µM in a solution containing 0.1 M ammonium acetate, 1 M guanidine-HCl and 2.5 mM reduced/0.25 mM oxidized glutathione (pH 8.0 with aqueous NH4OH) and stirred slowly at 4°C for 3 days. The folding reaction was monitored with RP-HPLC and the crude oxidized product was purified by preparative RP-HPLC with a C18 silica column. The purity of the synthetic PcTx1, GxTx1E and the Ala mutants was confirmed by analytical RP-HPLC and MALDI-TOF-MS measurements. The concentration of the toxin was determined by measuring absorbance at 280 nm using calculated extinction coefficients (Gill and von Hippel, 1989 (link)).
Gupta K., Zamanian M., Bae C., Milescu M., Krepkiy D., Tilley D.C., Sack J.T., Yarov-Yarovoy V., Kim J.I, & Swartz K.J. (2015). Tarantula toxins use common surfaces for interacting with Kv and ASIC ion channels. eLife, 4, e06774.
+ Open protocol
+ Expand
3

Soluble Aβ42 Peptide Preparation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Aβ42 and [F10, Y42]Aβ42 were synthesized using solid phase peptide synthesis and Fmoc chemistry on an Applied Biosystems model 433A peptide synthesizer (Foster City, CA, USA), as described previously [3 (link)]. Peptide lyophilizates (200 μ g of approximately 80% peptide by weight) were dissolved in 25 μ L of 60 mM sodium hydroxide (Fisher, Waltham, MA, USA) in water to increase solubility and decrease de novo peptide aggregation [29 (link)]. Immediately thereafter, 112.5 μ L of water and 112.5 μL of 22.2 mM sodium phosphate, pH 7.4, were added, and the solution was sonicated in an ultrasonic water bath (model 1510, Branson Ultrasonics Corp., Danbury, CT, USA) for 1 min at 22°C. The pH of the solution at this stage of peptide preparation is ≈11, which facilitates peptide manipulation by increasing solubility and preventing spontaneous self-association [29 (link)]. Protein concentration was determined at 22°C by UV absorbance (ε274=1280 cm−1M−1) using a 1 cm quartz cuvette (Hellma, Plainview, NY, USA) and a Beckman DU-640 spectrophotometer (Beckman Instruments, Fullerton, CA, USA). Peptide concentration was adjusted to 80 μ M using 10 mM sodium phosphate, pH 7.4. This yielded a final pH of 7.4.
Hayden E.Y., Conovaloff J.L., Mason A., Bitan G, & Teplow D.B. (2016). Preparation of pure populations of covalently stabilized amyloid β-protein oligomers of specific sizes. Analytical biochemistry, 518, 78-85.
+ Open protocol
+ Expand
4

Solid-Phase Peptide Synthesis Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
All peptides used in the evaluations of bioactivity were synthesized by solid-phase synthesis on an Applied Biosystems model 433A peptide synthesizer according to standard protocols, as previously reported32 (link). Purities of the synthetic peptides were greater than 95%.
Yang X., Wang Y., Zhang Y., Lee W.H, & Zhang Y. (2016). Rich diversity and potency of skin antioxidant peptides revealed a novel molecular basis for high-altitude adaptation of amphibians. Scientific Reports, 6, 19866.
+ Open protocol
+ Expand
5

Synthesis and Purification of Conformationally Restricted CCR5 Peptides

Check if the same lab product or an alternative is used in the 5 most similar protocols
Peptides were synthesized by the solid-phase Fmoc method (28 (link)) using an Applied Biosystems model 433A peptide synthesizer. After peptide assembly, resin-bound peptides were deprotected as previously described (29 (link)) and purified to greater than 95% purity by semipreparative reverse-phase high-performance liquid chromatography. To obtain conformationally restricted etherocyclic peptides, an extra cysteine was added at peptides covering ECL1 extracellular regions as previously described (19 (link)–21 (link)). All extracellular regions of CCR5 have been synthesized as shown in Table 1 and in particular amino-terminal region from aa 14 to 34 as it has been demonstrated to be immunogenic whereas peptide from 1 to 13 did not induce antibodies (20 (link), 21 (link)); ECL1 as constrained peptide, as its linear form did not elicit antibodies (19 (link)–21 (link)); ECL2 and ECL3 regions in their linear sequence.
Verna A.E., Franceschi V., Tebaldi G., Macchi F., Menozzi V., Pastori C., Lopalco L., Ottonello S., Cavirani S, & Donofrio G. (2017). Induction of Antihuman C–C Chemokine Receptor Type 5 Antibodies by a Bovine Herpesvirus Type-4 Based Vector. Frontiers in Immunology, 8, 1402.
+ Open protocol
+ Expand
6

Purification and Characterization of mAb#12 Fab Fragment

Check if the same lab product or an alternative is used in the 5 most similar protocols
Ascites containing mAb#12 IgG were produced by Harlan Bioproducts for Science (Indianapolis, IN), as previously described (13 (link)). A HiTrap Protein G column (GE Healthcare Life Sciences, Piscataway, NJ) was used to extract the IgG from the ascites solution. The Fab fragment of mAb#12 was prepared with the Fab preparation kit from Thermo Scientific (Rockford, IL) by following the manufacturer’s instructions. The Fab fragment was then further purified to homogeneity by a Mono S ion-exchange column (GE Healthcare Life Sciences). All peptides were chemically synthesized by the Core Laboratory of the Center for Biologics Evaluation and Research (CBER) at the US Food and Drug Administration (FDA), with an Applied Biosystems (Foster City, CA) model 433A peptide synthesizer. The mAb#12–epitope II complex was made by mixing the 26-mer peptide (421HINSTALNCNESLNTGWLAGLFYQHK446) with mAb#12 Fab (molar ratio of 4:1).
Deng L., Ma L., Virata M.L., Zhong L., Yan H., Zhao Z., Struble E., Feinstone S., Alter H, & Zhang P. (2014). Discrete conformations of epitope II on the hepatitis C virus E2 protein for antibody-mediated neutralization and nonneutralization. Proceedings of the National Academy of Sciences of the United States of America, 111(29), 10690-10695.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!