FLAMBE: A kinetic fluorescence polarization assay to study activation of monomeric BAX

Mohammed, Jarvier N.; Gelles, Jesse D.; Chipuk, Jerry E.

doi:10.1016/j.xpro.2022.101252

Cited by 4 publications

(6 citation statements)

References 10 publications

(28 reference statements)

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“…Negative experiences arising from learning during the pandemic has made tertiary education less attractive, and this pose as motivational drawbacks that hinder students’ decision to participate in tertiary education (Edge Research & HCM Strategists, 2022). Across the globe, similar trend is observed where 72.1% of 560,000 Malaysian school leavers did not wish to pursue further education, main causes cited were loss of interest in study, questioning the purpose of schooling, financial constraints, and the exam-centric system (Mohammed, 2022). With this, the current study aims to investigate the possible effects of negative emotions on college admission decision.…”

Section: Introductionmentioning

confidence: 69%

Emotion Matters: Understanding the Role of Achievement Emotions in High School Students’ Decision for Tertiary Education

Yim,

Sedhu,

Abdul-Rahim

2023

Journal of Psychoeducational Assessment

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Emotions play an integral role in learning. It is an effective response to a stimulus which can be experienced felt along a positive–negative continuum. Achievement emotions arise from academic activities, and can activate or de-activate educational motivation responses. Based on Control Value Theory, this study examined how negative achievement emotions impact high school students’ decision to pursue tertiary education, and what were the contributing factors of these emotions. One thousand five hundred and forty-seven high school students participated in the survey, and data were solicited via established inventories as well as an open ended question. Quantitatively, Partial Least Squares structural equation modelling found boredom and test anxiety to significantly lower students’ decision to pursue tertiary education. The precursors to negative achievement emotions stem from examinations and assessments, struggle in lessons, learning itself, parental expectations to excel, and mental health issues. Findings demonstrated the potential of emotions in influencing students’ value orientation with respect to tertiary education. As the study is based on a post-pandemic context, implications of findings are discussed in view of the underlying post-pandemic challenges accompanied by recommendation to stakeholders.

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Section: Introductionmentioning

confidence: 69%

Emotion Matters: Understanding the Role of Achievement Emotions in High School Students’ Decision for Tertiary Education

Yim,

Sedhu,

Abdul-Rahim

2023

Journal of Psychoeducational Assessment

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“…FLAMBE assays are performed as previously described (Gelles et al, 2022, Mohammed et al, 2022). Briefly, using a 96 well format and 100 μl total volume per condition, BAX, BIM BH3 , C8-BID, buffers, and fluorescently-labelled BAK BH3 peptide (BAK TAMRA ) are combined as indicated and analyzed for fluorescence (Excitation wavelength: 530 nm; Emission wavelength: 590 nm) using a multi-mode microplate reader fit with a polarization filter.…”

Section: Methodsmentioning

confidence: 99%

An optimized high-yield protocol for expression and purification of monomeric full-length BAX protein

Chen,

Gelles,

Mohammed

et al. 2023

Preprint

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Diverse developmental signals and pro-death stresses converge on regulation of the mitochondrial pathway of apoptosis. BAX, a pro-apoptotic BCL-2 effector, directly forms proteolipid pores in the outer mitochondrial member to activate the mitochondrial pathway of apoptosis. BAX is a viable pharmacological target for various human diseases, and increasing efforts have been made to study the molecular regulation of BAX and identify small molecules selectively targeting BAX. However, generating large quantities of monomeric and functionally-competent BAX has been challenging due to its aggregation-prone nature. Additionally, there is a lack of detailed and instructional protocols available for investigators who are not already familiar with recombinant BAX production. Here, we present a comprehensive high-yield protocol for expressing, purifying, and storing functional recombinant BAX protein. We utilize an intein-tagged BAX construct and employ a two-step chromatography strategy to capture and purify BAX, and provide example standard assays to observe BAX activation. We also highlight best practices for handling and storing BAX to effectively preserve its quality, shelf-life, and function.

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“…In each experiment, the BAK TAMRA control is shown in grey. A more detailed step-by-step protocol has been co-published in STAR Protocols ( Mohammed et al, 2022 ).…”

Section: Methodsmentioning

confidence: 99%

FLAMBE: A kinetic fluorescence polarization assay to study activation of monomeric BAX

Cited by 4 publications

References 10 publications

Emotion Matters: Understanding the Role of Achievement Emotions in High School Students’ Decision for Tertiary Education

Emotion Matters: Understanding the Role of Achievement Emotions in High School Students’ Decision for Tertiary Education

An optimized high-yield protocol for expression and purification of monomeric full-length BAX protein

A kinetic fluorescence polarization ligand assay for monitoring BAX early activation

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