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Lanthascreen eu anti gst

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LanthaScreen Eu‐anti‐GST is a fluorescence-based assay reagent used for the detection and quantification of glutathione S‐transferase (GST) fusion proteins. It utilizes a europium-labeled anti‐GST antibody to provide a time-resolved fluorescence signal in the presence of the target GST fusion protein. The core function of this product is to enable researchers to monitor and analyze GST‐tagged proteins in a variety of applications, such as high‐throughput screening and biochemical assays.

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Lab products found in correlation

Proxiplate, by PerkinElmer (2 mentions) Gst hdac10, by Thermo Fisher Scientific (2 mentions) Tubastatin alexa647 tracer, by Thermo Fisher Scientific (2 mentions)

2 protocols using lanthascreen eu anti gst

1

HDAC10 Inhibition Assay via TR-FRET

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The FRET‐assay was performed in white 384‐well ProxiPlates (PerkinElmer). Reagents were diluted in assay buffer (50 mM HEPES pH 8.0, 150 mM NaCl, 10 mM MgCl2, 1 mM EGTA, 0.01 % Brij‐35). Final assay volume (10 μL) contains 3 nM GST‐HDAC10 (Life Technologies), 30 nM Tubastatin‐Alexa647‐Tracer,[32] and 0.5 nM LanthaScreen Eu‐anti‐GST (Life Technologies). Compound stocks (10 mM DMSO) were diluted in assay buffer. An 11‐fold 1 : 3‐serial dilution of test compounds (1 μL) were presented in the plate and complemented by 9 μL of assay mix. After incubation (1 h, room temperature) TR‐FRET was measured with EnVision plate reader (Ex: 3 flashes of the TRF‐europiumlaser; Em: 620 and 665 nm (665/620 nm ratio). Inhibition was calculated by using negative control (2 % DMSO) and positive control (20 μM vorinostat). Dose‐response curves were fitted in ActivityBase (IDBS) using a four‐parameter logistic model and IC50‐values were calculated.[32]
Herp D., Ridinger J., Robaa D., Shinsky S.A., Schmidtkunz K., Yesiloglu T.Z., Bayer T., Steimbach R.R., Herbst‐Gervasoni C.J., Merz A., Romier C., Sehr P., Gunkel N., Miller A.K., Christianson D.W., Oehme I., Sippl W, & Jung M. (2022). First Fluorescent Acetylspermidine Deacetylation Assay for HDAC10 Identifies Selective Inhibitors with Cellular Target Engagement. Chembiochem, 23(14), e202200180.
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2

HDAC10 Enzyme Inhibition Assay

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The FRET-assay was performed in white 384-well ProxiPlates (PerkinElmer). Reagents were diluted in assay buffer (50 mM HEPES pH 8.0, 150 mM NaCl, 10 mM MgCl2, 1 mM EGTA, 0.01% Brij-35). Final assay volume (10 μL) contains 3 nM GST-HDAC10 (Life Technologies), 30 nM Tubastatin-Alexa647-Tracer[32 (link)], and 0.5 nM LanthaScreen Eu-anti-GST (Life Technologies). Compound stocks (10 mM DMSO) were diluted in assay buffer. An 11-fold 1:3-serial dilution of test compounds (1 μL) were presented in the plate and complemented by 9 μL of assay mix. After incubation (1 h, room temperature) TR-FRET was measured with EnVision plate reader (Ex: 3 flashes of the TRF-europiumlaser; Em: 620 and 665 nm (665/620 nm ratio). Inhibition was calculated by using negative control (2 % DMSO) and positive control (20 μM vorinostat). Dose-response curves were fitted in ActivityBase (IDBS) using a four-parameter logistic model and IC50-values were calculated.[32 (link)]
Herp D., Ridinger J., Robaa D., Shinsky S.A., Schmidtkunz K., Yesiloglu T.Z., Bayer T., Steimbach R.R., Herbst‐Gervasoni C.J., Merz A., Romier C., Sehr P., Gunkel N., Miller A.K., Christianson D.W., Oehme I., Sippl W, & Jung M. (2022). First Fluorescent Acetylspermidine Deacetylation Assay for HDAC10 Identifies Selective Inhibitors with Cellular Target Engagement. Chembiochem, 23(14), e202200180.
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