In Vitro Cell Based Assay

1. IN-VITRO CELL BASED ASSAY- CORE ASSAY LIST 2011

2. CORE ASSAY LIST 2011
THERAPEUTIC AREA – IMMUNOMODULATION
Turn Minimum
Project
REF# Assay Principle Around Compound Standard Format
Type
Time Required
Test model: Determine effect of mitogens/inhibitors on cell
proliferation in murine splenocytes/Human PBMC. Useful for lead Up to 3 compounds at
Cell
CA01 optimization of proliferation/ anti-proliferation compounds. 4 Weeks 10 mg 5 concentrations tested
Proliferation
Assay format: Incubations in 48-well plates. MTT read out. in quadruplicate
Data format: WordTM report with data available in an ExcelTM file.
Test model: Determine effect of compounds on (un-stimulated)
Nitric Oxide release from murine macrophages. Useful for lead
Up to 4 compounds at
Mediator optimization for pro-inflammatory compounds and potency
CA02 4 Weeks 10 mg 5 concentrations tested
release determination.
in triplicate
Assay format: Incubations in 96-well plates. Colorimetric.
Data format: WordTM report with data available in an ExcelTM file.
Test model: Determine effect of compounds on (stimulated)
cytokine release (IL-2) from murine splenocytes. Useful for lead
Up to 3 compounds at
Mediator optimization for pro-inflammatory compounds and potency
CA03 4 Weeks 10 mg 5 concentrations tested
release determination.
in quadruplicate
Assay format: Incubations in 48-well plates. ELISA.
Data format: WordTM report with data available in an ExcelTM file.
Test model: Determine effect of compounds on (stimulated)
cytokine release (IL-3) from murine splenocytes. Useful for lead
Up to 3 compounds at
Mediator optimization for pro-inflammatory compounds and potency
CA04 4 Weeks 10 mg 5 concentrations tested
release determination.
in quadruplicate
Assay format: Incubations in 48-well plates. ELISA.
Data format: WordTM report with data available in an ExcelTM file.
R & D Centre: Plot # 5B, 19th K.M Stone, Hosur Road, Bangalore – 560 100, India.
Tel: 91-080-7832265, 7833092, 7834872, Fax: 91-080-7834369, E-mail: cvc@naturalremedy.com

3. THERAPEUTIC AREA – IMMUNOMODULATION
Turn Minimum
Project
REF# Assay Principle Around Compound Standard Format
Type
Time Required
Test model: Determine effect of compounds on (stimulated)
cytokine release (IL-4) from murine splenocytes. Useful for lead
Up to 3 compounds at
Mediator optimization for Anti-inflammatory compounds and potency
CA05 4 Weeks 10 mg 5 concentrations tested
release determination.
in quadruplicate
Assay format: Incubations in 48-well plates. ELISA.
Data format: WordTM report with data available in an ExcelTM file.
Test model: Determine effect of compounds on (stimulated)
cytokine release (IL-5) from murine splenocytes. Useful for lead
Up to 3 compounds at
Mediator optimization for pro-inflammatory compounds and potency
CA06 4 Weeks 10 mg 5 concentrations tested
release determination.
in quadruplicate
Assay format: Incubations in 48-well plates. ELISA.
Data format: WordTM report with data available in an ExcelTM file.
Test model: Determine effect of compounds on (stimulated)
cytokine release (IL-6) from murine splenocytes. Useful for lead
Up to 3 compounds at
Mediator optimization for Anti-inflammatory compounds and potency
CA07 4 Weeks 10 mg 5 concentrations tested
release determination.
in quadruplicate
Assay format: Incubations in 48-well plates. ELISA.
Data format: WordTM report with data available in an ExcelTM file.
Test model: Determine effect of compounds on (stimulated)
cytokine release (IL-10) from murine splenocytes. Useful for lead
Up to 3 compounds at
Mediator optimization for Anti-inflammatory compounds and potency
CA08 4 Weeks 10 mg 5 concentrations tested
release determination.
in quadruplicate
Assay format: Incubations in 48-well plates. ELISA.
Data format: WordTM report with data available in an ExcelTM file.
R & D Centre: Plot # 5B, 19th K.M Stone, Hosur Road, Bangalore – 560 100, India.
Tel: 91-080-7832265, 7833092, 7834872, Fax: 91-080-7834369, E-mail: cvc@naturalremedy.com

4. THERAPEUTIC AREA – IMMUNOMODULATION
Turn Minimum
Project
REF# Assay Principle Around Compound Standard Format
Type
Time Required
Test model: Determine effect of compounds on (stimulated)
cytokine release (IL-12) from murine splenocytes. Useful for lead
Up to 3 compounds at
Mediator optimization for Anti-inflammatory compounds and potency
CA09 4 Weeks 10 mg 5 concentrations tested
release determination.
in quadruplicate
Assay format: Incubations in 48-well plates. ELISA.
Data format: WordTM report with data available in an ExcelTM file.
Test model: Determine effect of compounds on (stimulated)
cytokine release (IFN-γ) from murine splenocytes. Useful for lead
Up to 3 compounds at
Mediator optimization for pro-inflammatory compounds and potency
CA10 4 Weeks 10 mg 5 concentrations tested
release determination.
in quadruplicate
Assay format: Incubations in 48-well plates. ELISA.
Data format: WordTM report with data available in an ExcelTM file.
R & D Centre: Plot # 5B, 19th K.M Stone, Hosur Road, Bangalore – 560 100, India.
Tel: 91-080-7832265, 7833092, 7834872, Fax: 91-080-7834369, E-mail: cvc@naturalremedy.com

5. THERAPEUTIC AREA – ALLERGY/PAIN AND INFLAMMATION
Turn Minimum
Project
REF# Assay Principle Around Compound Standard Format
Type
Time Required
Test model: Determine effect of compounds on (stimulated) Nitric
Oxide release from murine macrophages. Useful for lead
Up to 4 compounds at
Mediator optimization for Anti-inflammatory compounds and potency
CA11 4 Weeks 10 mg 5 concentrations tested
release determination.
in triplicate
Assay format: Incubations in 96-well plates. Colorimetric.
Data format: WordTM report with data available in an ExcelTM file.
Test model: Determine effect of compounds on (stimulated)
Prostaglandin release (PGE2) from murine macrophages. Useful
Up to 4 compounds at
Mediator for lead optimization for Anti-inflammatory compounds and
CA12 4 Weeks 10 mg 5 concentrations tested
release potency determination.
in triplicate
Assay format: Incubations in 96-well plates. HTRF.
Data format: WordTM report with data available in an ExcelTM file.
Test model: Determine effect of compounds on (stimulated)
Thromboxane release (TXB2) from Human pro-myelocytic
Up to 4 compounds at
Mediator leukemia cells. Useful for lead optimization for Anti-inflammatory
CA13 4 Weeks 10 mg 5 concentrations tested
release compounds and potency determination.
in triplicate
Assay format: Incubations in 96-well plates. ELISA.
Data format: WordTM report with data available in an ExcelTM file.
Test model: Determine effect of compounds on (stimulated)
Leukotriene release (LTB4) from Human pro-myelocytic leukemia
Up to 4 compounds at
Mediator cells. Useful for lead optimization for Anti-inflammatory
CA14 4 Weeks 10 mg 5 concentrations tested
release compounds and potency determination.
in triplicate
Assay format: Incubations in 96-well plates. HTRF.
Data format: WordTM report with data available in an ExcelTM file.
R & D Centre: Plot # 5B, 19th K.M Stone, Hosur Road, Bangalore – 560 100, India.
Tel: 91-080-7832265, 7833092, 7834872, Fax: 91-080-7834369, E-mail: cvc@naturalremedy.com

6. THERAPEUTIC AREA – ALLERGY/PAIN AND INFLAMMATION
Turn Minimum
Project
REF# Assay Principle Around Compound Standard Format
Type
Time Required
Test model: Determine effect of compounds on (stimulated)
cytokine release (IL-1β) from murine macrophages. Useful for lead
Up to 4 compounds at
Mediator optimization for Anti-inflammatory compounds and potency
CA15 4 Weeks 10 mg 5 concentrations tested
release determination.
in triplicate
Assay format: Incubations in 96-well plates. ELISA.
Data format: WordTM report with data available in an ExcelTM file.
Test model: Determine effect of compounds on (stimulated)
Histamine release from rat basophilic leukemia cells. Useful for
Up to 4 compounds at
Mediator lead optimization for Anti-allergic compounds and potency
CA16 4 Weeks 10 mg 5 concentrations tested
release determination.
in triplicate
Assay format: Incubations in 96-well plates. HTRF.
Data format: WordTM report with data available in an ExcelTM file.
Test model: Determine effect of compounds on (stimulated)
cytokine release (MCP-1) from murine peritoneal macrophages.
Up to 4 compounds at
Mediator Useful for lead optimization for Anti-inflammatory compounds and
CA17 4 Weeks 10 mg 5 concentrations tested
release potency determination.
in triplicate
Assay format: Incubations in 96-well plates. ELISA.
Data format: WordTM report with data available in an ExcelTM file.
R & D Centre: Plot # 5B, 19th K.M Stone, Hosur Road, Bangalore – 560 100, India.
Tel: 91-080-7832265, 7833092, 7834872, Fax: 91-080-7834369, E-mail: cvc@naturalremedy.com

7. THERAPEUTIC AREA – IN VITRO TOXICITY- HEPATOPROTECTION
Turn Minimum
Project
REF# Assay Principle Around Compound Standard Format
Type
Time Required
Test model: Measurement of compound toxicity in hepatocytes
against t-BH using viability (MTT) endpoints. Assess compound
Up to 4 compounds at
In vitro safety before proceeding to clinical development.
CA18 4 Weeks 10 mg 5 concentrations tested
Toxicity Assay format: Incubations in 96-well plates. In vitro toxicity
in triplicate
profiling.
Data format: Word™ report with data available in an Excel™ file.
Test model: Measurement of compound toxicity in hepatocytes
against diazinon (OPC) using viability (MTT) endpoints. Assess
Up to 4 compounds at
In vitro compound safety before proceeding to clinical development.
CA19 4 Weeks 10 mg 5 concentrations tested
Toxicity Assay format: Incubations in 96-well plates. In vitro toxicity
in triplicate
profiling.
Data format: Word™ report with data available in an Excel™ file.
Test model: Measurement of compound toxicity in hepatocytes
against Aldrin (OC) using viability (MTT) endpoints. Assess
Up to 4 compounds at
In vitro compound safety before proceeding to clinical development.
CA19 4 Weeks 10 mg 5 concentrations tested
Toxicity Assay format: Incubations in 96-well plates. In vitro toxicity
in triplicate
profiling.
Data format: Word™ report with data available in an Excel™ file.
Test model: Measurement of compound toxicity in hepatocytes
against Aflatoxin (AFB1) using viability (MTT) endpoints. Assess
Up to 4 compounds at
In vitro compound safety before proceeding to clinical development.
CA20 4 Weeks 10 mg 5 concentrations tested
Toxicity Assay format: Incubations in 96-well plates. In vitro toxicity
in triplicate
profiling.
Data format: Word™ report with data available in an Excel™ file.
R & D Centre: Plot # 5B, 19th K.M Stone, Hosur Road, Bangalore – 560 100, India.
Tel: 91-080-7832265, 7833092, 7834872, Fax: 91-080-7834369, E-mail: cvc@naturalremedy.com

8. THERAPEUTIC AREA – IN VITRO TOXICITY- HEPATOPROTECTION
Turn Minimum
Project
REF# Assay Principle Around Compound Standard Format
Type
Time Required
Test model: Measurement of compound toxicity in hepatocytes
against Galactosamine using viability (MTT) endpoints. Assess
Up to 4 compounds at
In vitro compound safety before proceeding to clinical development.
CA21 4 Weeks 10 mg 5 concentrations tested
Toxicity Assay format: Incubations in 96-well plates. In vitro toxicity
in triplicate
profiling.
Data format: Word™ report with data available in an Excel™ file.
THERAPEUTIC AREA – COSMETOLOGY
Turn Minimum
Project
REF# Assay Principle Around Compound Standard Format
Type
Time Required
Test model: Determine effect of compounds on (un-stimulated)
collagen release from primary human dermal fibroblast. Useful for Up to 4 compounds at
CA22 Anti-wrinkle lead optimization for cosmetology ingredient. 4 Weeks 10 mg 5 concentrations tested
Assay format: Incubations in 96-well plates. Colorimetric. in triplicate
Data format: WordTM report with data available in an ExcelTM file.
Test model: Determine effect of compounds on (un-stimulated)
Melanin release from mouse melanoma cells. Useful for lead Up to 4 compounds at
Skin
CA23 optimization for cosmetology ingredient. 4 Weeks 10 mg 5 concentrations tested
whitening
Assay format: Incubations in 96-well plates. Colorimetric. in triplicate
Data format: WordTM report with data available in an ExcelTM file.
R & D Centre: Plot # 5B, 19th K.M Stone, Hosur Road, Bangalore – 560 100, India.
Tel: 91-080-7832265, 7833092, 7834872, Fax: 91-080-7834369, E-mail: cvc@naturalremedy.com

9. THERAPEUTIC AREA – DIABETES/OBESITY
Turn Minimum
REF# Project Type Assay Principle Around Compound Standard Format
Time Required
Test Model: Measurement of induction of glucose stimulated
Insulin secretion in rat insulinoma cells. This approach is useful for
assessing the effectiveness of compounds under development for Up to 4 compounds at
CA24 GSIS Assay the treatment of type 1 diabetes. 4 Weeks 10 mg 5 concentrations tested
Assay Format: Incubations in 96-well plates. HTRF. in triplicate
Data Format: WordTM report with data available in an ExcelTM
file.
Test model: Determine effect of compounds on (stimulated)
Lipid
Adipocyte differentiation in Mouse embryonic fibroblast . Useful Up to 4 compounds at
Metabolism
CA25 for lead optimization for inhibitors of adipogenesis. 4 Weeks 10 mg 5 concentrations tested
Assays
Assay format: Incubations in 96-well plates. Colorimetric. in triplicate
(Adipogenesis)
Data format: WordTM report with data available in an ExcelTM file.
Test model: Determine effect of compounds on (stimulated)
Lipid
glycerol release in Mouse embryonic fibroblast . Useful for loead Up to 4 compounds at
Metabolism
CA26 identification for inhibitors of adipolysis. 4 Weeks 10 mg 5 concentrations tested
Assays
Assay format: Incubations in 96-well plates. Colorimetric. in triplicate
(Adipolysis)
Data format: WordTM report with data available in an ExcelTM file.
R & D Centre: Plot # 5B, 19th K.M Stone, Hosur Road, Bangalore – 560 100, India.
Tel: 91-080-7832265, 7833092, 7834872, Fax: 91-080-7834369, E-mail: cvc@naturalremedy.com

10. THERAPEUTIC AREA – ONCOLOGY
Turn Minimum
REF# Project Type Assay Principle Around Compound Standard Format
Time Required
Test Model: Determine effect of compounds on cell proliferation
in human breast adenocarcinoma cells. Useful for lead optimization
Anti Up to 4 compounds at
of anti-cancer compound screening.
CA27 proliferative 4 Weeks 10 mg 5 concentrations tested
assay
Assay Format: Incubations in 96-well plates. Colorimetric.
in triplicate
Data Format: WordTM report with data available in an ExcelTM
file.
THERAPEUTIC AREA – GENERAL HEALTH/ HEART DISEASE
Turn Minimum
REF# Project Type Assay Principle Around Compound Standard Format
Time Required
Test Model: Determine the cellular antioxidant potential of
compounds in Human hepatocellular liver carcinoma cells. Useful
cost-effective method to evaluate the antioxidant activity of foods, Up to 4 compounds at
Antioxidant
CA28 dietary supplements, phytochemicals and extracts.. 4 Weeks 10 mg 5 concentrations tested
assay
Assay Format: Incubations in 96-well plates. Colorimetric. in triplicate
Data Format: WordTM report with data available in an ExcelTM
file.
R & D Centre: Plot # 5B, 19th K.M Stone, Hosur Road, Bangalore – 560 100, India.
Tel: 91-080-7832265, 7833092, 7834872, Fax: 91-080-7834369, E-mail: cvc@naturalremedy.com

11. THERAPEUTIC AREA – IN VITRO GENOTOXICITY
Turn Minimum
REF# Project Type Assay Principle Around Compound Standard Format
Time Required
Test Model:. Determine the mutageneic potential of compounds
using 5 starins of S.Typhimurium with and without S9 fractions.
Up to 1 compounds at
Mutagenesis Quick assay to estimate the carcinogenic potential of a compound
CA29 4 Weeks 10 mg 6 concentrations tested
assay Assay Format: Incubations in 384-well plates. Hightroughput.
in duplicate.
Data Format: WordTM report with data available in an ExcelTM
file.
Test model: Determine effect of compounds on the formation of
small membrane bound DNA fragments i.e. micronuclei in the
cytoplasm of interphase cells using Chinese hamster cells. Useful Up to 1 compounds at
Mutagenesis
CA30 for early identification of compounds likely to exhibit genotoxic 15 Weeks 10 mg 3 concentrations tested
assay
liability. in duplicate.
Assay format: Incubations in 90mm petri plates. Manual.
Data format: WordTM report with data available in an ExcelTM file.
Test model: Determine effect of compounds on induction of
structural changes in chromosomes using Chinese hamster cells.
Up to 1 compounds at
Cytogenetic Useful to identify agents that cause structural chromosome
CA31 15 Weeks 10 mg 3 concentrations tested
Assays aberration in cultured mammalian cells.
in duplicate.
Assay format: Incubations in 90mm petri plates. Manual.
Data format: WordTM report with data available in an ExcelTM file.
R & D Centre: Plot # 5B, 19th K.M Stone, Hosur Road, Bangalore – 560 100, India.
Tel: 91-080-7832265, 7833092, 7834872, Fax: 91-080-7834369, E-mail: cvc@naturalremedy.com