Alexa fluor 532 c5 maleimide

Manufactured by Thermo Fisher Scientific

Alexa Fluor 532 C5 maleimide is a fluorescent dye used for labeling proteins and other biomolecules. It has an absorption maximum at 532 nm and an emission maximum at 554 nm. The maleimide functional group allows for covalent attachment to thiol-containing compounds.

Automatically generated - may contain errors

Lab products found in correlation

Pd 10 desalting column, by GE Healthcare (1 mentions) Zeba spin column, by Thermo Fisher Scientific (1 mentions) Nickel 2 chloride hexahydrate, by Merck Group (1 mentions) Sulfo cyanine5 maleimide, by Lumiprobe (1 mentions) Dtssp, by Thermo Fisher Scientific (1 mentions) Bafilomycin a1, by InvivoGen (1 mentions) L leucyl l leucine methyl ester hydrochloride, by Cayman Chemical (1 mentions) Heparin sodium salt from porcine intestinal mucosa, by Merck Group (1 mentions)

4 protocols using alexa fluor 532 c5 maleimide

Fluorescent Labeling of Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Gun4 was reduced in 10 mM DTT for 10 min at room temperature before buffer exchange into 20 mM NaH₂PO₄ and 150 mM NaCl, pH 7.4, using a Zeba Spin column (Thermo Scientific) and then labelled with a 10-fold excess of Alexa Fluor 532 C₅ maleimide (Life Technologies), dissolved in dry DMF, for 2 h at room temperature while being protected from the light. The reaction was quenched by the addition of 10 mM DTT for 15 min.
Wild-type (WT) ChlH and ΔN159ChlH were desalted into PBS titrated to pH 8.3 with 0.2 M pH 9 sodium bicarbonate, using a Zeba Spin column. Protein was labelled with a 3-fold excess of Atto 488 NHS ester dissolved in dry DMF for 1 h at room temperature while being protected from light. The reaction was quenched with the addition of 10 mM Tris/HCl (pH 7.4) for 15 min.
Proteins were separated from excess dye by using a PD-10 desalting column (GE Healthcare) equilibrated in 50 mM Tricine/NaOH, 200 mM NaCl and 0.3 M glycerol.

Adams N.B., Marklew C.J., Qian P., Brindley A.A., Davison P.A., Bullough P.A, & Hunter C.N. (2014). Structural and functional consequences of removing the N-terminal domain from the magnesium chelatase ChlH subunit of Thermosynechococcus elongatus. Biochemical Journal, 464(Pt 3), 315-322.

+ Open protocol

+ Expand

Fluorescent Labeling of Tau and Kinesin Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

3RS Tau protein was incubated with a 10 fold molar excess of dithiothreitol (DTT) for two hours at room temperature and DTT was removed using a 2 mL 7K MWCO Zeba™ Spin Desalting Column (Pierce, Rockford, IL). Tau was then incubated in a 10 fold molar excess of Alexa Fluor 532-C5 maleimide (Life Technologies, Grand Island, NY) for an additional 2 hours at room temperature, and excess fluorophore was removed using a second desalting column. Labeling efficiency of Tau was determined by comparing the concentration of fluorophore to protein. Tau concentration was determined with the BCA assay and the dye concentration was determined using an extinction coefficient of 81,000 M⁻¹*cm⁻¹ at 531 nm (Alexa Fluor 532) in a Beckman DU^® 640 spectrophotometer (Beckman Coulter, Brea, CA). The 3RS-isoform was labeled at C233 and the labeling efficiency was determined to be 85%. Rigor kinesin protein was labeled similarly except it was incubated with DTT for two hours on ice followed with Alexa Fluor 532-C5 maleimide at 4°C overnight. Rigor kinesin was labeled at any of four possible solvent-exposed cysteines (C7, C66, C169 and C296)^{36 (link)} and the labeling efficiency was determined to be 95%.

Hoeprich G.J., Mickolajczyk K.J., Nelson S.R., Hancock W.O, & Berger C.L. (2017). The Axonal Transport Motor Kinesin-2 Navigates Microtubule Obstacles via Protofilament Switching. Traffic (Copenhagen, Denmark), 18(5), 304-314.

+ Open protocol

+ Expand

Labeling Proteins with Fluorescent Dyes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Nickel(ii) chloride hexahydrate was obtained from Sigma-Aldrich. DTSSP (3,3′-dithiobis(sulfosuccinimidylpropionate)), Alexa Fluor™ 532 C5 maleimide and Anti-His₆-tag mouse monoclonal antibody conjugated with HRP (MA1-21315-HRP) were purchased from ThermoFisher Scientific. Sulfo-Cyanine5 maleimide was purchased from Lumiprobe.

Vervoort D.F., Pretto C, & van Hest J.C. (2020). Insight into N-terminal localization and dynamics of engineered virus-like particles. RSC Advances, 10(64), 38774-38781.

+ Open protocol

+ Expand

Bafilomycin A1 and LLOME Labeling Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Bafilomycin A1 was purchased from InvivoGen. L-Leucyl-L-Leucine methyl ester (hydrochloride) (LLOME) was purchased from Cayman chemicals. Both compounds were dissolved in DMSO and small aliquots were stored at −20 °C until further use. For non-treated samples, DMSO alone was used as a control. Alexa Fluor 532 C5 maleimide and Alexa Fluor 647 N-hydroxysuccinimide (NHS) ester were purchased from Thermo Fischer Scientific and freshly dissolved in DMSO before protein labeling. Heparin sodium salt from porcine intestinal mucosa was purchased from Merck (H3393).

Yuste-Checa P., Trinkaus V.A., Riera-Tur I., Imamoglu R., Schaller T.F., Wang H., Dudanova I., Hipp M.S., Bracher A, & Hartl F.U. (2021). The extracellular chaperone Clusterin enhances Tau aggregate seeding in a cellular model. Nature Communications, 12, 4863.

+ 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?

Revolutionizing how scientists
search and build protocols!