scholarly journals Spesifitas dan Sensitifitas Antibodi Anti eRF3 Ragi Saccharomyces cerevisia

2007 ◽  
Vol 1 (1) ◽  
Author(s):  
Sandra Hermanto
Keyword(s):  
Western Blot ◽  
Crude Extract ◽  
Release Factor ◽  
Wild Type

Protein eRF3 (eukaryotic release factor-3) merupakan salah satu protein yang berperan padaproses terminasi translasi. Protein ini bersama-sama dengan eRF1 (eukaryotic release factor-1) saling berinteraksi membentuk kompleks release factor dalam memediasi pelepasan rantaipolipeptida dari ribosom. Untuk memahami mekanisme terminasi translasi dalam sistemeukariot telah dilakukan studi struktur fungsi eRF1 yang dilanjutkan dengan studi interaksi invitro eRF1 mutan dan eRF1 wild type dengan eRF3. Namun demikian, hasil deteksi dari studiinteraksi in vitro sulit terdeteksi secara kuantitatif. Untuk dapat mengkuantisasi pita-pitaeRF3 hasil studi interaksi in vitro diperlukan antibodi anti eRF3. Konstruksi antibodi antieRF3 telah dilakukan, tetapi antibodi ini belum terkarakterisasi dengan baik. Tahapanselanjutnya dilakukan analisa Western blot dengan cara mengukur tingkat spesifitas dansensitifitas antibodi anti eRF3 terhadap protein eRF3. Spesifitas antibodi ditentukanberdasarkan kemampuan antibodi ini dalam mengenali epitop protein eRF3 dari berbagaiprotein yang terdapat pada crude extract ragi, sedangkan sensitifitasnya ditentukan melaluivariasi jumlah antigen (eRF3) yang berinteraksi dengan antibodi tersebut. Hasil analisaWestern blot menunjukkan spesifitas antibodi anti eRF3 masih relatif baik dimana antibodiini mampu mengenali epitop protein eRF3 yang ditandai dengan munculnya pita tunggal(76,6 kDa) setelah antibodi ini direaksikan dengan crude extract ragi yang mengandungprotein eRF3. Sensitifitas antibodi ini juga relatif tinggi, karena antibodi ini mampumendeteksi protein eRF3 hingga jumlah yang relatif rendah (0,77 ng). Namun demikianantibodi ini belum cukup mampu mendeteksi protein eRF3 yang secara alamiah terdapat padacrude extract ragi. Hal ini kemungkinan besar disebabkan karena level ekspresi eRF3 dalamsel ragi yang relatif rendah jika dibandingkan dengan protein ribosom.

scholarly journals HGG-26. H3G34V MUTATION AFFECTS GENOMIC H3K36 METHYLATION IN PEDIATRIC GLIOMA

2020 ◽  
Vol 22 (Supplement_3) ◽  
pp. iii348-iii348
Author(s):  
Tina Huang ◽  
Andrea Piunti ◽  
Elizabeth Bartom ◽  
Jin Qi ◽  
Rintaro Hashizume ◽  
...  
Keyword(s):  
Western Blot ◽  
Allelic Frequency ◽  
Glioma Cell Line ◽  
Fluorescent Imaging ◽  
Wild Type ◽  
Gene Expressions ◽  
Pediatric Glioma ◽  
Genes Encoding ◽  
Normal Human

Abstract BACKGROUND Histone H3.3 mutation (H3F3A) occurs in 50% of cortical pediatric high-grade gliomas. This mutation replaces glycine 34 with arginine or valine (G34R/V), impairing SETD2 activity (H3K36-specific trimethyltransferase), resulting in reduced H3K36me on H3G34V nucleosomes relative to wild-type. This contributes to genomic instability and drives distinct gene expressions associated with tumorigenesis. However, it is not known if this differential H3K36me3 enrichment is due to H3G34V mutant protein alone. Therefore, we set to elucidate the effect of H3G34V on genomic H3K36me3 enrichment in vitro. METHODS Doxycycline-inducible short hairpin RNA (shRNA) against H3F3A was delivered via lentivirus to established H3G34V mutant pediatric glioma cell line KNS42, and H3G34V introduced into H3.3 wild type normal human astrocytes (NHA). Transfections were confirmed by western blot, fluorescent imaging, and flow cytometry, with resulting H3.3WT and H3K36me3 expression determined by western blot. H3.3WT, H3K36me3, and H3G34V ChIP-Seq was performed to evaluate genomic enrichment. RESULTS Complete knockdown of H3G34V was achieved with DOX-induced shRNA, with no change in total H3.3, suggesting disproportionate allelic frequency of genes encoding H3.3 (H3F3A and H3F3B). Modest increase in H3K36me3 occurred after H3F3A-knockdown from KNS42, suggesting H3G34V alone impacts observed H3K36me3 levels. Distinct H3K36me3 genomic enrichment was observed with H3G34V knock-in. CONCLUSIONS We demonstrate that DOX-inducible knockdown of H3F3A in an H3G34V mutant pediatric glioma cells and H3G34V mutation transduction in wild-type astrocytes affects H3K36me3 expression. Further evaluation by ChIP-Seq analysis for restoration of wild-type genomic H3K36me3 enrichment patterns with H3G34V knockdown, and mutant H3K36me3 patterns with H3G34V transduction, is currently underway.

scholarly journals Increased Association of Deamidated αA-N101D with Lens Membrane of Transgenic αAN101D vs. Wild Type αA Mice: Potential Effects on Intracellular Ionic Imbalance and Membrane Disorganization

2020 ◽  
Author(s):  
Om Srivast ◽  
Kiran Srivast ◽  
Roy Joseph ◽  
Landon Wilson
Keyword(s):  
Epithelial Cells ◽  
Western Blot ◽  
Immunogold Labeling ◽  
Human Lens ◽  
Membrane Association ◽  
Related Protein ◽  
Wild Type ◽  
Age Related ◽  
Ionic Imbalance

Abstract We have generated two mouse models, in one by inserting the human lens αAN101D transgene in CRYαAN101D mice, and in the other by inserting human wild-type αA-transgene in CRYαAWT mice. The CRYαAN101D mice developed cortical cataract at about 7-months of age relative to CRYαAWT mice. The objective of the study was to determine the following relative changes in the lenses of CRYαAN101D- vs. CRYαAWT mice: age-related changes with specific emphasis on protein insolubilization, relative membrane-association of αAN101D vs. WTαA proteins, and changes in intracellular ionic imbalance and membrane organization. Methods: Lenses of varying ages from CRYαAWT and CRYαAN101D mice were compared for an age-related protein insolubilization. The relative lens membrane-association of the αAN101D- and WTαA proteins in the two types of mice was determined by immunohistochemical-, immunogold-labeling-, and western blot analyses. The relative levels of membrane-binding of recombinant αAN101D- and WTαA proteins was determined by an in vitro assay, and the levels of intracellular Ca2+ uptake and Na, K-ATPase mRNA were determined in the cultured epithelial cells from lenses of the two types of mice.Results: Compared to the lenses of CRYαAWT, the lenses of CRYαAN101D mice exhibited: (A) An increase in age-related protein insolubilization beginning at about 4-months of age. (B) A greater lens membrane-association of αAN101D- relative to WTαA protein during immunogold-labeling- and western blot analyses, including relatively a greater membrane swelling in the CRYαAN101D lenses. (C) During in vitro assay, the greater levels of binding αAN101D- relative to WTαA protein to membranes was observed. (D) The 75% lower level of Na, K-ATPase mRNA but 1.5X greater Ca2+ uptake were observed in cultured lens epithelial cells of CRYαAN101D- than those of CRYαAWT mice. Conclusions: The results show that an increased lens membrane association of αAN101D--relative WTαA protein in CRYαAN101D mice than CRYαAWT mice occurs, which causes intracellular ionic imbalance, and in turn, membrane swelling that potentially leads to cortical opacity.

scholarly journals Increased Association of Deamidated αA-N101D with Lens Membrane of Transgenic αAN101D vs. Wild Type αA Mice: Potential Effects on Intracellular Ionic Imbalance and Membrane Disorganization 

2020 ◽  
Author(s):  
Om Srivast ◽  
Kiran Srivast ◽  
Roy Joseph ◽  
Landon Wilson
Keyword(s):  
Western Blot ◽  
In Vitro Assay ◽  
Immunogold Labeling ◽  
Membrane Association ◽  
Related Protein ◽  
Wild Type ◽  
Vitro Assay ◽  
Age Related ◽  
Ionic Imbalance

Abstract We have generated two mouse models, in one by inserting the human lens αAN101D transgene in CRYαA N101D mice, and in the other by inserting human wild-type αA-transgene in CRYαA WT mice. The CRYαA N101D mice developed cortical cataract at about 7-months of age relative to CRYαA WT mice. The objective of the study was to determine the following relative changes in the lenses of CRYαA N101D - vs. CRYαA WT mice: age-related changes with specific emphasis on protein insolubilization, relative membrane-association of αA N101D vs. WTαA proteins, and changes in intracellular ionic imbalance and membrane organization. Methods: Lenses of varying ages from CRYαA WT and CRYαA N101D mice were compared for an age-related protein insolubilization. The relative lens membrane-association of the αAN101D- and WTαA proteins in the two types of mice was determined by immunohistochemical-, immunogold-labeling-, and western blot analyses. The relative levels of membrane-binding of recombinant αA N101D - and WTαA proteins was determined by an in vitro assay, and the levels of intracellular Ca 2+ uptake and Na, K-ATPase mRNA were determined in the cultured epithelial cells from lenses of the two types of mice. Results: Compared to the lenses of CRYαA WT , the lenses of CRYαA N101D mice exhibited: (A) An increase in age-related protein insolubilization beginning at about 4-months of age. (B) A greater lens membrane-association of αAN101D- relative to WTαA protein during immunogold-labeling- and western blot analyses, including relatively a greater membrane swelling in the CRYαA N101D lenses. (C) During in vitro assay, the greater levels of binding αAN101D- relative to WTαA protein to membranes was observed. (D) The 75% lower level of Na, K-ATPase mRNA but 1.5X greater Ca 2+ uptake was observed in cultured lens epithelial cells of CRYαA N101D- than those of CRYαA WT mice. Conclusions: The results show that an increased lens membrane association of αA N101D - - relative WTαA protein in CRYαA N101D mice than CRYαA WT mice occurs, which causes intracellular ionic imbalance, and in turn, membrane swelling that potentially leads to cortical opacity.

scholarly journals Increased Association of Deamidated αA-N101D with Lens Membrane of Transgenic αAN101D vs. Wild Type αA Mice: Potential Effects on Intracellular Ionic Imbalance and Membrane Disorganization

2019 ◽  
Author(s):  
Om Srivast ◽  
Kiran Srivast ◽  
Roy Joseph ◽  
Landon Wilson
Keyword(s):  
Western Blot ◽  
In Vitro Assay ◽  
Immunogold Labeling ◽  
Membrane Association ◽  
Related Protein ◽  
Wild Type ◽  
Vitro Assay ◽  
Age Related ◽  
Ionic Imbalance

Abstract Background: We have generated mouse models by inserting the human lens αA-N101D transgene in CRYAAN101D mice, and human wild-type αA-transgene in CRYAAWT mice. The CRYAAN101D mice developed cortical cataract at about 7-months of age relative to CRYAAWT mice. The objective of the study was to determine the following relative changes in the lenses of CRYAAN101D- vs. CRYAAWT mice: age-related changes with specific emphasis on protein insolubilization, relative membrane-association of αAN101D vs. WTαA, and changes intracellular ionic imbalance and membrane organization. Methods: Lenses from CRYAAWT and CRYAAN101D mice were compared for an age-related protein insolubilization. The relative lens membrane-association of the αAN101D and WTαA in the two types of mice was determined by immunohistochemical-, immunogold-labeling-, and western blot analyses. The relative levels of membrane-binding of recombinant αAN101D and WTαA was determined by an in vitro assay, and the levels of intracellular Ca 2+ uptake and Na, K-ATPase mRNA were determined in the cultured epithelial cells of the two types lenses. Results: Compared to the lenses of CRYAAWT, the lenses of CRYAAN101D mice exhibited: (A) An increase in age-related protein insolubilization beginning at about 4-months of age. (B) A greater lens membrane-association of αAN101D relative to WTαA during immunogold-labeling- and western blot analyses, including relatively a greater membrane swelling in the CRYAAN101D lenses. . (C) During in vitro assay, the greater levels of binding αAN101D to membranes relative to WTαA was observed. (D) The 75% lower level of Na,K-ATPase mRNA but 1.5X greater Ca 2+ uptake were observed in cultured lens epithelial cells of CRYAAN101D than those of CRYAAWT mice. Conclusions: The results show that an increased lens membrane association of αAN101D relative WTαA in CRYAAN101D mice than CRYAAWT mice, which causes intracellular ionic imbalance, and in turn membrane swelling leading to cortical opacity.

scholarly journals PDTM-31. INVESTIGATING THE EFFECT OF H3G34V MUTATION ON DISTINCT GENOMIC H3K36 METHYLATION PATTERNS IN PEDIATRIC GLIOMA

2019 ◽  
Vol 21 (Supplement_6) ◽  
pp. vi194-vi194
Author(s):  
Tina Huang ◽  
Andrea Piunti ◽  
Patrick Ozark ◽  
Elizabeth Barton ◽  
Jin Qi ◽  
...  
Keyword(s):  
Western Blot ◽  
Cell Line ◽  
Glioma Cell ◽  
Gene Expression Signature ◽  
Allelic Frequency ◽  
Glioma Cell Line ◽  
Fluorescent Imaging ◽  
Wild Type ◽  
Pediatric Glioma

Abstract BACKGROUND Histone H3.3 mutation (H3F3A) occurs in 50% of cortical pediatric high-grade gliomas (pHGGs). This mutation replaces glycine 34 with arginine or valine (G34R/V), impairing SETD2 activity (H3K36-specific trimethyltransferase). Consequently, reduced H3K36me is observed on H3G34V nucleosomes relative to wild-type, contributing to genomic instability and driving a distinct gene expression signature associated with tumorigenesis. However, it is not known if this differential H3K36me3 enrichment is due to H3G34V mutant protein alone. Therefore, we set to elucidate the effect of H3G34V mutation in pediatric glioma on genomic H3K36me3 enrichment in vitro. METHODS Doxycycline-inducible short hairpin RNA (shRNA) against H3F3A was delivered via lentivirus to established H3G34V mutant pediatric glioma cell line KNS42, and H3G34V introduced into H3.3 wild type normal human astrocytes (NHA). Transfections were confirmed by western blot, fluorescent imaging, and flow cytometry, with resulting H3.3WT and H3K36me3 expression determined by western blot. H3.3WT, H3K36me3, and H3G34V ChIP-Seq was performed to evaluate effects of H3G34V mutation on genomic enrichment patterns. RESULTS Complete knockdown of H3G34V was achieved with DOX-induced shRNA, with no change in total H3.3, suggesting disproportionate allelic frequency of genes encoding H3.3 (H3F3A and H3F3B). Modest increase in H3K36me3 expression occurred after H3F3A-knockdown from KNS42, suggesting H3G34V alone impacts observed H3K36me3 levels. H3G34V knock-in to NHAs was verified via western blot. Distinct H3K36me3 genomic enrichment was observed with H3G34V on ChIP-Seq. CONCLUSIONS We demonstrate that DOX-inducible knockdown of H3F3A in an H3G34V mutant pediatric glioma cell line, and H3G34V mutation transduction in wild-type NHAs, directly impacts H3K36me3 expression level. Further evaluation by ChIP-Seq analysis for restoration of wild-type genomic H3K36me3 enrichment patterns with H3G34V knockdown, and mutant H3K36me3 patterns with H3G34V transduction, is currently underway.

scholarly journals Increased Association of Deamidated αA-N101D with Lens Membrane of Transgenic αAN101D vs. Wild Type αA Mice: Potential Effects on Intracellular Ionic Imbalance and Membrane Disorganization 

2020 ◽  
Author(s):  
Om Srivast ◽  
Kiran Srivast ◽  
Roy Joseph ◽  
Landon Wilson
Keyword(s):  
Epithelial Cells ◽  
Western Blot ◽  
Immunogold Labeling ◽  
Human Lens ◽  
Membrane Association ◽  
Related Protein ◽  
Wild Type ◽  
Age Related ◽  
Ionic Imbalance

Abstract We have generated two mouse models, in one by inserting the human lens αAN101D transgene in CRYαAN101D mice, and in the other by inserting human wild-type αA-transgene in CRYαAWT mice. The CRYαAN101D mice developed cortical cataract at about 7-months of age relative to CRYαAWT mice. The objective of the study was to determine the following relative changes in the lenses of CRYαAN101D- vs. CRYαAWT mice: age-related changes with specific emphasis on protein insolubilization, relative membrane-association of αAN101D vs. WTαA proteins, and changes in intracellular ionic imbalance and membrane organization. Methods: Lenses of varying ages from CRYαAWT and CRYαAN101D mice were compared for an age-related protein insolubilization. The relative lens membrane-association of the αAN101D- and WTαA proteins in the two types of mice was determined by immunohistochemical-, immunogold-labeling-, and western blot analyses. The relative levels of membrane-binding of recombinant αAN101D- and WTαA proteins was determined by an in vitro assay, and the levels of intracellular Ca2+ uptake and Na, K-ATPase mRNA were determined in the cultured epithelial cells from lenses of the two types of mice.Results: Compared to the lenses of CRYαAWT, the lenses of CRYαAN101D mice exhibited: (A) An increase in age-related protein insolubilization beginning at about 4-months of age. (B) A greater lens membrane-association of αAN101D- relative to WTαA protein during immunogold-labeling- and western blot analyses, including relatively a greater membrane swelling in the CRYαAN101D lenses. (C) During in vitro assay, the greater levels of binding αAN101D- relative to WTαA protein to membranes was observed. (D) The 75% lower level of Na, K-ATPase mRNA but 1.5X greater Ca2+ uptake were observed in cultured lens epithelial cells of CRYαAN101D- than those of CRYαAWT mice. Conclusions: The results show that an increased lens membrane association of αAN101D--relative WTαA protein in CRYαAN101D mice than CRYαAWT mice occurs, which causes intracellular ionic imbalance, and in turn, membrane swelling that potentially leads to cortical opacity.

scholarly journals Increased Association of Deamidated αA-N101D with Lens membrane of transgenic αAN101D vs. wild type αA mice: potential effects on intracellular ionic imbalance and membrane disorganization

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Om Srivastava ◽  
Kiran Srivastava ◽  
Roy Joseph ◽  
Landon Wilson
Keyword(s):  
Western Blot ◽  
In Vitro Assay ◽  
Immunogold Labeling ◽  
Membrane Association ◽  
Related Protein ◽  
Wild Type ◽  
Vitro Assay ◽  
Age Related ◽  
Ionic Imbalance

Abstract We have generated two mouse models, in one by inserting the human lens αAN101D transgene in CRYαAN101D mice, and in the other by inserting human wild-type αA-transgene in CRYαAWT mice. The CRYαAN101D mice developed cortical cataract at about 7-months of age relative to CRYαAWT mice. The objective of the study was to determine the following relative changes in the lenses of CRYαAN101D- vs. CRYαAWT mice: age-related changes with specific emphasis on protein insolubilization, relative membrane-association of αAN101D vs. WTαA proteins, and changes in intracellular ionic imbalance and membrane organization. Methods Lenses of varying ages from CRYαAWT and CRYαAN101D mice were compared for an age-related protein insolubilization. The relative lens membrane-association of the αAN101D- and WTαA proteins in the two types of mice was determined by immunohistochemical-, immunogold-labeling-, and western blot analyses. The relative levels of membrane-binding of recombinant αAN101D- and WTαA proteins was determined by an in vitro assay, and the levels of intracellular Ca2+ uptake and Na, K-ATPase mRNA were determined in the cultured epithelial cells from lenses of the two types of mice. Results Compared to the lenses of CRYαAWT, the lenses of CRYαAN101D mice exhibited: (A) An increase in age-related protein insolubilization beginning at about 4-months of age. (B) A greater lens membrane-association of αAN101D- relative to WTαA protein during immunogold-labeling- and western blot analyses, including relatively a greater membrane swelling in the CRYαAN101D lenses. (C) During in vitro assay, the greater levels of binding αAN101D- relative to WTαA protein to membranes was observed. (D) The 75% lower level of Na, K-ATPase mRNA but 1.5X greater Ca2+ uptake were observed in cultured lens epithelial cells of CRYαAN101D- than those of CRYαAWT mice. Conclusions The results show that an increased lens membrane association of αAN101D-−relative WTαA protein in CRYαAN101D mice than CRYαAWT mice occurs, which causes intracellular ionic imbalance, and in turn, membrane swelling that potentially leads to cortical opacity.

scholarly journals Increased Association of Deamidated αA-N101D with Lens Membrane of Transgenic αAN101D vs. Wild Type αA Mice: Potential Effects on Intracellular Ionic Imbalance and Membrane Disorganization

2020 ◽  
Author(s):  
Om Srivast ◽  
Kiran Srivast ◽  
Roy Joseph ◽  
Landon Wilson
Keyword(s):  
Epithelial Cells ◽  
Western Blot ◽  
Immunogold Labeling ◽  
Human Lens ◽  
Membrane Association ◽  
Related Protein ◽  
Wild Type ◽  
Age Related ◽  
Ionic Imbalance

Abstract We have generated two mouse models, in one by inserting the human lens αAN101D transgene in CRYαAN101D mice, and in the other by inserting human wild-type αA-transgene in CRYαAWT mice. The CRYαAN101D mice developed cortical cataract at about 7-months of age relative to CRYαAWT mice. The objective of the study was to determine the following relative changes in the lenses of CRYαAN101D- vs. CRYαAWT mice: age-related changes with specific emphasis on protein insolubilization, relative membrane-association of αAN101D vs. WTαA proteins, and changes in intracellular ionic imbalance and membrane organization. Methods: Lenses of varying ages from CRYαAWT and CRYαAN101D mice were compared for an age-related protein insolubilization. The relative lens membrane-association of the αAN101D- and WTαA proteins in the two types of mice was determined by immunohistochemical-, immunogold-labeling-, and western blot analyses. The relative levels of membrane-binding of recombinant αAN101D- and WTαA proteins was determined by an in vitro assay, and the levels of intracellular Ca2+ uptake and Na, K-ATPase mRNA were determined in the cultured epithelial cells from lenses of the two types of mice.Results: Compared to the lenses of CRYαAWT, the lenses of CRYαAN101D mice exhibited: (A) An increase in age-related protein insolubilization beginning at about 4-months of age. (B) A greater lens membrane-association of αAN101D- relative to WTαA protein during immunogold-labeling- and western blot analyses, including relatively a greater membrane swelling in the CRYαAN101D lenses. (C) During in vitro assay, the greater levels of binding αAN101D- relative to WTαA protein to membranes was observed. (D) The 75% lower level of Na, K-ATPase mRNA but 1.5X greater Ca2+ uptake was observed in cultured lens epithelial cells of CRYαAN101D- than those of CRYαAWT mice. Conclusions: The results show that an increased lens membrane association of αAN101D--relative WTαA protein in CRYαAN101D mice than CRYαAWT mice occurs, which causes intracellular ionic imbalance, and in turn, membrane swelling that potentially leads to cortical opacity.

Genotoxicity evaluation in vitro by comet assay of compound and crude extract obtained from P. Pubescens Benth.

Planta Medica ◽  
2015 ◽  
Vol 81 (11) ◽  
Author(s):  
VHS Souza ◽  
A Paula OHohne ◽  
R Grando ◽  
N de Cassia de Almeida Queiroz ◽  
GM Pastore ◽  
...  
Keyword(s):  
Comet Assay ◽  
Crude Extract

Phytochemical analysis and salivary amylase inhibition activities of Carica papaya leaf and Garcinia mangostana pericarp extracts and partially purified fractions

2017 ◽  
Vol 6 (1) ◽  
pp. 34
Author(s):  
Michael Russelle Alvarez ◽  
Paolo Robert Bueno ◽  
Raymond Oliver Cruz ◽  
Richard Macapulay ◽  
Francis Jayson Vallesfin ◽  
...  
Keyword(s):  
Crude Extract ◽  
Carica Papaya ◽  
Uv Light ◽  
Digestive Enzyme ◽  
Phytochemical Analysis ◽  
Phytochemical Screening ◽  
Salivary Amylase ◽  
Garcinia Mangostana ◽  
Leaf Extracts

Plant-derived digestive enzyme inhibitors particularly those targeted to carbohydrate metabolism has been the focus of recent studies as natural supplements for weight control and diabetes. The present study explores the salivary amylase inhibition activity of Garcinia mangostana (Linn.) pericarp extracts and Carica papaya (Linn.) leaf extracts and fractions, as well as perform phytochemical screening and quantification, and thin layer – and high performance liquid chromatographic profiling. ­Results show that crude extracts and purified fractions were able to inhibit salivary amylase, with C. papaya fraction 1 being the most active at 30.89% inhibition. Phytochemical screening of all extracts tested ­positive for tannins, glycosides, phenolics, flavonoids and alkaloids. Quantification of phenolics showed that extracts contained high levels of phenolics, with C. papaya crude extract having the highest content with 219.0±12.7 mg GAE/g extract followed by G. mangostana crude extract with 247.1±18.0 mg GAE/g extract. Quantification of total flavonoids also showed C. papaya crude extract to contain the highest content with 55.12±0.679 mg QE/g extract. All extracts contained negligible alkaloid content, though. HPLC and TLC profiling showed several peaks and bands, when viewed in 210 nm and UV light, respectively. These results demonstrate in vitro the salivary amylase inhibitory activity of both plants and their potential as antidiabetic drug candidates; however, further studies need to be done, like isolation and structure elucidation of active components and toxicity assays. Keywords: Amylase inhibition, phytochemical quantification, Carica papaya, Garcinia mangostana

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