Cat. No.: HY-50847

ZSTK474

18 Cited Publications

Customer Review

Based on 18 publication(s) in Google Scholar

ZSTK474 is an ATP-competitive pan-class I PI3K inhibitor with IC₅₀s of 16 nM, 44 nM, 4.6 nM and 49 nM for PΙ3Κα, PI3Kβ, PI3Kδ and PI3Kγ, respectively.

For research use only. We do not sell to patients.

Purity: 99.71%
CAS No.: 475110-96-4
Formula: C₁₉H₂₁F₂N₇O₂
Molecular Weight:417.41

Storage:

Powder -20°C, 3 years , 4°C, 2 years ; In solvent -80°C, 2 years , -20°C, 1 year

ZSTK474 Chemical Structure

CAS No.: 475110-96-4

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Fluorescent Dye

Strong Fluorescence Signal / Multi-Disciplinary Applications / High Fluorescence Specificity

Biological Activity
- Description
- IC₅₀ & Target
- Cellular Effect
- In Vitro
- In Vivo
- Clinical Trial
Chemical Information
Publications (18)
Solvent & Solubility
- In Vitro
- In Vivo
Protocol
Purity & Documentation
References
Help & FAQs
Classifications
Bioz

Biological Activity

Description

ZSTK474 is an ATP-competitive pan-class I PI3K inhibitor with IC₅₀s of 16 nM, 44 nM, 4.6 nM and 49 nM for PΙ3Κα, PI3Kβ, PI3Kδ and PI3Kγ, respectively.

IC₅₀ & Target^[2]

PI3Kδ

4.6 nM (IC₅₀)

PI3Kα

16 nM (IC₅₀)

PI3Kβ

44 nM (IC₅₀)

PI3Kγ

49 nM (IC₅₀)

Autophagy

All PI3K Isoforms :

Cellular Effect

Cell Line	Type	Value	Description	References
A549	IC₅₀	17 nM Compound: ZSTK474	Inhibition of PI3K p110 beta in human A549 cell lysate preincubated with compound for 5 mins followed by [gamma-32p]-ATP addition measured after 20 mins by immunoprecipitation based liquid scintillation spectroscopic analysis Inhibition of PI3K p110 beta in human A549 cell lysate preincubated with compound for 5 mins followed by [gamma-32p]-ATP addition measured after 20 mins by immunoprecipitation based liquid scintillation spectroscopic analysis	[PMID: 16622124]
A549	IC₅₀	37 nM Compound: ZSTK474	Inhibition of PI3K in human A549 cell lysate preincubated with compound for 5 mins followed by [gamma-32p]-ATP addition measured after 20 mins by immunoprecipitation based liquid scintillation spectroscopic analysis Inhibition of PI3K in human A549 cell lysate preincubated with compound for 5 mins followed by [gamma-32p]-ATP addition measured after 20 mins by immunoprecipitation based liquid scintillation spectroscopic analysis	[PMID: 16622124]
A549	IC₅₀	53 nM Compound: ZSTK474	Inhibition of PI3K p110 gamma in human A549 cell lysate preincubated with compound for 5 mins followed by [gamma-32p]-ATP addition measured after 20 mins by immunoprecipitation based liquid scintillation spectroscopic analysis Inhibition of PI3K p110 gamma in human A549 cell lysate preincubated with compound for 5 mins followed by [gamma-32p]-ATP addition measured after 20 mins by immunoprecipitation based liquid scintillation spectroscopic analysis	[PMID: 16622124]
A549	IC₅₀	6 nM Compound: ZSTK474	Inhibition of PI3K p110 delta in human A549 cell lysate preincubated with compound for 5 mins followed by [gamma-32p]-ATP addition measured after 20 mins by immunoprecipitation based liquid scintillation spectroscopic analysis Inhibition of PI3K p110 delta in human A549 cell lysate preincubated with compound for 5 mins followed by [gamma-32p]-ATP addition measured after 20 mins by immunoprecipitation based liquid scintillation spectroscopic analysis	[PMID: 16622124]
HCT-116	IC₅₀	0.71 μM Compound: IV; ZSTK-474	Antiproliferative activity against human HCT-116 cells assessed as inhibition of cell proliferation incubated for 96 hrs by MTT assay Antiproliferative activity against human HCT-116 cells assessed as inhibition of cell proliferation incubated for 96 hrs by MTT assay	[PMID: 37736169]
HCT-116	IC₅₀	1.31 μM Compound: ZSTK474	Antiproliferative activity against human HCT116 cells harboring PI3Kalpha H1047R mutant after 72 hrs by CCK8 assay Antiproliferative activity against human HCT116 cells harboring PI3Kalpha H1047R mutant after 72 hrs by CCK8 assay	[PMID: 30071408]
HCT-116	IC₅₀	1.31 μM Compound: ZSTK474	Antiproliferative activity against human HCT116 cells after 72 hrs by CCK-8 assay Antiproliferative activity against human HCT116 cells after 72 hrs by CCK-8 assay	[PMID: 30245395]
HCT-116	IC₅₀	78 nM Compound: 1, ZSTK474	Inhibition of PIK3CA H1047R mutant-mediated cell signaling in human HCT116 cells expressing PTEN assessed as inhibition of insulin-induced pAkt/PKB phosphorylation at Thr308 treated for 15 mins before insulin challenge measured after 5 mins by immunoblott Inhibition of PIK3CA H1047R mutant-mediated cell signaling in human HCT116 cells expressing PTEN assessed as inhibition of insulin-induced pAkt/PKB phosphorylation at Thr308 treated for 15 mins before insulin challenge measured after 5 mins by immunoblott	[PMID: 21882832]
HCT-116	IC₅₀	97 nM Compound: 1, ZSTK474	Inhibition of PIK3CA H1047R mutant-mediated cell signaling in human HCT116 cells expressing PTEN assessed as inhibition of insulin-induced pAkt/PKB phosphorylation at Ser473 treated for 15 mins before insulin challenge measured after 5 mins by immunoblott Inhibition of PIK3CA H1047R mutant-mediated cell signaling in human HCT116 cells expressing PTEN assessed as inhibition of insulin-induced pAkt/PKB phosphorylation at Ser473 treated for 15 mins before insulin challenge measured after 5 mins by immunoblott	[PMID: 21882832]
HT-29	GI₅₀	300 nM Compound: ZSTK474	Antiproliferative activity against human HT-29 cells incubated for 3 days by WST8 assay Antiproliferative activity against human HT-29 cells incubated for 3 days by WST8 assay	[PMID: 33284613]
LNCaP	IC₅₀	210 nM Compound: 6	Antiproliferative activity against human LNCAP cells after 3 days by MTS assay Antiproliferative activity against human LNCAP cells after 3 days by MTS assay	[PMID: 20227881]
MCF7	IC₅₀	5.27 μM Compound: ZSTK474	Antiproliferative activity against human MCF7 cells after 72 hrs by CCK8 assay Antiproliferative activity against human MCF7 cells after 72 hrs by CCK8 assay	[PMID: 30071408]
MCF7	IC₅₀	5.27 μM Compound: ZSTK474	Antiproliferative activity against human MCF7 cells after 72 hrs by CCK-8 assay Antiproliferative activity against human MCF7 cells after 72 hrs by CCK-8 assay	[PMID: 30245395]
MDA-MB-468	EC₅₀	3.2 μM Compound: ZSTK474	Cytotoxicity against PTEN-deficient human MDA-MB-468 cells assessed as inhibition of cell growth after 48 hrs by Cell Titer 96 assay Cytotoxicity against PTEN-deficient human MDA-MB-468 cells assessed as inhibition of cell growth after 48 hrs by Cell Titer 96 assay	[PMID: 23795239]
Sf21	IC₅₀	0.003 μM Compound: 4a, ZSTK474	Inhibition of PI3Kdelta (unknown origin) expressed in SF21 cells using phosphatidylinositol as substrate after 60 mins by Kinase-Glo assay Inhibition of PI3Kdelta (unknown origin) expressed in SF21 cells using phosphatidylinositol as substrate after 60 mins by Kinase-Glo assay	[PMID: 23265896]
Sf21	IC₅₀	0.006 μM Compound: 4a, ZSTK474	Inhibition of PI3Kalpha (unknown origin) expressed in SF21 cells using phosphatidylinositol as substrate after 60 mins by Kinase-Glo assay Inhibition of PI3Kalpha (unknown origin) expressed in SF21 cells using phosphatidylinositol as substrate after 60 mins by Kinase-Glo assay	[PMID: 23265896]
Sf21	IC₅₀	0.006 μM Compound: 4a, ZSTK474	Inhibition of PI3Kbeta (unknown origin) expressed in SF21 cells using phosphatidylinositol as substrate after 60 mins by Kinase-Glo assay Inhibition of PI3Kbeta (unknown origin) expressed in SF21 cells using phosphatidylinositol as substrate after 60 mins by Kinase-Glo assay	[PMID: 23265896]
Sf21	IC₅₀	0.038 μM Compound: 4a, ZSTK474	Inhibition of PI3Kgamma (unknown origin) expressed in SF21 cells using phosphatidylinositol as substrate after 60 mins by Kinase-Glo assay Inhibition of PI3Kgamma (unknown origin) expressed in SF21 cells using phosphatidylinositol as substrate after 60 mins by Kinase-Glo assay	[PMID: 23265896]
Sf21	IC₅₀	6 nM Compound: ZSTK-47	Inhibition of PI3Kbeta (unknown origin) expressed in Escherichia coli-infected fall armyworm sf21 cells co-expressing p85 by kinase-glo luminescence assay Inhibition of PI3Kbeta (unknown origin) expressed in Escherichia coli-infected fall armyworm sf21 cells co-expressing p85 by kinase-glo luminescence assay	[PMID: 27387421]
U-87MG ATCC	IC₅₀	0.13 μM Compound: IV; ZSTK-474	Antiproliferative activity against human U-87 MG cells assessed as inhibition of cell proliferation incubated for 96 hrs by MTT assay Antiproliferative activity against human U-87 MG cells assessed as inhibition of cell proliferation incubated for 96 hrs by MTT assay	[PMID: 37736169]
U-87MG ATCC	IC₅₀	111 nM Compound: 1, ZSTK474	Inhibition of PIK3CA-mediated cell signaling in PTEN-deficient human U87MG cells assessed as inhibition of insulin-induced pAkt/PKB phosphorylation at Ser473 treated for 15 mins before insulin challenge measured after 5 mins by immunoblotting Inhibition of PIK3CA-mediated cell signaling in PTEN-deficient human U87MG cells assessed as inhibition of insulin-induced pAkt/PKB phosphorylation at Ser473 treated for 15 mins before insulin challenge measured after 5 mins by immunoblotting	[PMID: 21882832]
U-87MG ATCC	IC₅₀	32 nM Compound: 1, ZSTK474	Inhibition of PIK3CA-mediated cell signaling in PTEN-deficient human U87MG cells assessed as inhibition of insulin-induced pAkt/PKB phosphorylation at Thr308 treated for 15 mins before insulin challenge measured after 5 mins by immunoblotting Inhibition of PIK3CA-mediated cell signaling in PTEN-deficient human U87MG cells assessed as inhibition of insulin-induced pAkt/PKB phosphorylation at Thr308 treated for 15 mins before insulin challenge measured after 5 mins by immunoblotting	[PMID: 21882832]

In Vitro

Lineweaver-Burk plot analysis revealed that ZSTK474 inhibits all four PI3K isoforms in an ATP-competitive manner. The K_i values determined for the four PI3K isoforms showed that ZSTK474 inhibited the PI3Kδ isoform most effectively with a K_i of 1.8 nM, whereas the other isoforms are inhibited with 4-10-fold higher K_i values. Therefore, ZSTK474 should be regarded as a pan-PI3K inhibitor. We also determined the IC₅₀ values for inhibiting the four PI3K isoforms with ZSTK474 and LY294002. The IC₅₀ values of ZSTK474 (16, 44, 4.6 and 49 nM for PI3Kα, PI3Kβ, PI3Kδ and PI3Kγ, respectively) are shown to be consistent with the K_i values (6.7, 10.4, 1.8 and 11.7 nM for PI3Kα, PI3Kβ, PI3Kδ and PI3Kγ, respectively), which further supported the idea that ZSTK474 inhibits PI3Kδ most potently. Even at a concentration of 100 μM, ZSTK474 inhibits mTOR activity rather weakly^[1].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

In Vivo

In mice subjected to MCAO, treatment with ZSTK474 is tested at dosages of 50, 100, 200, and 300 mg/kg. Since the 200 mg/kg dose produces significant improvement and no obvious toxic effects (P<0.01), mice are treated with ZSTK474 at a dose of 200 mg/kg/day daily for three post-MCAO days during the remaining experiments of this study. Neurological function is examined in mice suffered from MCAO followed by 24, 48, and 72 h of reperfusion. In the ZSTK474 group, neurological function scores are significantly better than the control group except the corner test^[2].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Clinical Trial

NCT Number	Sponsor	Condition	Start Date	Phase A549
NCT01329991	Plexxikon\|		2011-05	PHASE1

Chemical Information

CAS No. 475110-96-4
Appearance Solid
Molecular Weight 417.41
Formula C₁₉H₂₁F₂N₇O₂
Color White to off-white
SMILES

FC(C1=NC2=C(N1C3=NC(N4CCOCC4)=NC(N5CCOCC5)=N3)C=CC=C2)F
Shipping

Room temperature in continental US; may vary elsewhere.

Storage

Powder	-20°C	3 years
	4°C	2 years
In solvent	-80°C	2 years
	-20°C	1 year

Publications (18)

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Sci Transl Med

PP2A inhibition is a druggable MEK inhibitor resistance mechanism in KRAS-mutant lung cancer cells. [Abstract]

2018 Jul 18;10(450):eaaq1093. PMID: 30021885
J Exp Clin Cancer Res

Prolonged inhibition of class I PI3K promotes liver cancer stem cell expansion by augmenting SGK3/GSK-3β/β-catenin signalling. [Abstract]

2018 Jun 25;37(1):122. PMID: 29940988

ZSTK474 purchased from MedChemExpress. Usage Cited in: J Exp Clin Cancer Res. 2018 Jun 25;37(1):122. [Abstract]

MHCC97H cells are treated with either LY294002 (2.5, 5,10, 20, 40 μM) or ZSTK474 (0.5, 1, 2.5, 5, 10 μM) for 24 h. DMSO is used as the control. Cell lysates are subjected to western blot analysis with the indicated antibodies.
Cell Death Dis

YIF1A activates mTORC1 signaling to promote cellular senescence. [Abstract]

2026 Jun 23. PMID: 42336842
Clin Cancer Res

Upregulation of IGF1R by mutant RAS in leukemia and potentiation of RAS signaling inhibitors by small-molecule inhibition of IGF1R. [Abstract]

2014 Nov 1;20(21):5483-95. PMID: 25186968
Neurotherapeutics

Salidroside ameliorates experimental autoimmune neuritis by dually modulating neuroinflammation and immune homeostasis via PI3K/AKT signaling. [Abstract]

2025 Nov 7:e00769. PMID: 41206312
Mol Metab

The lysosomal LAMTOR / Ragulator complex is essential for nutrient homeostasis in brown adipose tissue. [Abstract]

2023 May:71:101705. PMID: 36907508
J Endocrinol

GLP-1 signaling is required for improvement of glucose tolerance by osteocalcin. [Abstract]

2020 Feb;244(2):285-296. PMID: 31693486
Molecules

In Vitro and in Vivo Activity of mTOR Kinase and PI3K Inhibitors Against Leishmania donovani and Trypanosoma brucei. [Abstract]

2020 Apr 23;25(8):1980. PMID: 32340370
BMC Cancer

MARCH1 negatively regulates TBK1-mTOR signaling pathway by ubiquitinating TBK1. [Abstract]

2024 Jul 26;24(1):902. PMID: 39061024
J Biol Chem

Kinome-wide selectivity profiling of ATP-competitive mammalian target of rapamycin (mTOR) inhibitors and characterization of their binding kinetics. [Abstract]

2012 Mar 23;287(13):9742-9752. PMID: 22223645
Gene

Birch pollen-induced signatures in dendritic cells are maintained upon additional cytomegalovirus exposure. [Abstract]

2024 Nov 15:927:148649. PMID: 38852697
Exp Ther Med

mTORC1 is a key regulator that mediates OGD- and TGFβ1-induced myofibroblast transformation and chondroitin-4-sulfate expression in cardiac fibroblasts. [Abstract]

2022 Jun;23(6):413. PMID: 35601064
Meat and Muscle Biology

2026.
bioRxiv

2024 September 07.
Université de Lausanne

2024 Feb 7.
Bioengineered

The phosphoinositide 3-kinase inhibitor ZSTK474 increases the susceptibility of osteosarcoma cells to oncolytic vesicular stomatitis virus VSVΔ51 via aggravating endoplasmic reticulum stress. [Abstract]

2021 Dec;12(2):11847-11857. PMID: 34720036
bioRxiv

2020 Mar.
Research Square Preprint

2020 Feb.

Solvent & Solubility

In Vitro:

DMSO : 33.33 mg/mL (79.85 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

View the Complete Stock Solution Preparation Table

Preparing Stock Solutions

View the Complete Stock Solution Preparation Table

Concentration Solvent Mass

1 mg

5 mg

10 mg

Concentration

Solvent

Mass

1 mM

2.3957 mL

11.9786 mL

23.9573 mL

5 mM

0.4791 mL

2.3957 mL

4.7915 mL

10 mM

0.2396 mL

1.1979 mL

2.3957 mL

Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.

Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.

Molarity Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

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Molecular Weight Molecular Weight

Dilution Calculator

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In Vivo:

The following protocol is derived from the literature and is for reference only. It is recommended to first try a small sample.

Protocol 1

ZSTK474 is suspended in 5% hydroxypropyl cellulose^[3].

In Vivo Dissolution Calculator

Please enter the basic information of animal experiments:

Dosage Dosage
mg/kg
Animal Weight (Per Animal) Animal Weight (Per Animal)
g
Dosing Volume (Per Animal) Dosing Volume (Per Animal)
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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.

Working solution concentration: 0.22 mg/mL

Protocol

Kinase Assay^[1]

The linear phase of each kinetic reaction is defined at the respective enzyme amount (0.05, 0.1, 0.12 and 1 µg/mL for PI3Kα, PI3β, PI3δ and PI3γ, respectively) and reaction time (20 min). PI3K activity is assayed at various concentrations of ATP (5, 10, 25, 50, 100 µM) in the presence of increasing concentrations of ZSTK474. A Lineweaver-Burk plot is developed by plotting 1/v (the inverse of v, where v is obtained by subtracting the HTRF signal of the kinase test sample from the HTRF signal of the minus-enzyme control) versus 1/[ATP] (the inverse of the ATP concentration). For the minus-enzyme control, PIP2 is incubated with ATP in the absence of kinase. To determine the K_i value (inhibition constant) of ZSTK474 for each PI3K isoform, the slope of the respective Lineweaver-Burk plot is replotted against the ZSTK474 concentration. The K_i values are calculated by analysis using GraphPad Prism 4^[1].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Animal Administration^[2]

Mice^[2]
Mice are randomly assigned to receive different doses of ZSTK474 (50, 100, 200, and 300 mg/kg) to determine the optimum dose; in our experiment, the optimum dose is 200 mg/kg. Then mice are randomly assigned to one of three groups: a sham-operated group (phosphate-buffered saline, PBS); a control group (MCAO+PBS); a ZSTK474-treated group (MCAO+ZSTK474). In the ZSTK474-treated group, the mice are given the optimum dose of 200 mg/kg ZSTK474. In the sham-operated group and control group, mice are given an equivalent volume of PBS. All mice receive that same dose daily via oral gavage beginning at 6 h after the onset of focal ischemia and continuing for two more days, i.e., for a total of 3 days.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Purity & Documentation

Select Batch:

Purity: 99.98%

Assay: 99.98%

ee.: 99.98%

Data Sheet (288 KB)
SDS (393 KB)
- English - EN (393 KB)
- Français - FR (393 KB)
- Deutsch - DE (393 KB)
- Norwegian - NO (393 KB)
- Español - ES (393 KB)
- Swedish - SV (393 KB)
- Italian - IT (393 KB)
- Korean - KR (393 KB)
- Portuguese - PT (393 KB)
COA (2659 KB)
Elemental Analysis Report (2659 KB)
HNMR (2659 KB)
CNMR (2659 KB)
RP-HPLC (2659 KB)
NP-HPLC (2659 KB)
MS (2659 KB)
LCMS (2659 KB)
IR (2659 KB)
GC (2659 KB)
OR (2659 KB)
SDS-PAGE (2659 KB)
SEC-HPLC (2659 KB)
Other Report (2659 KB)
Handling Instructions (2659 KB)

References

[1]. Kong D, et al. ZSTK474 is an ATP-competitive inhibitor of class I phosphatidylinositol 3 kinase isoforms. Cancer Sci, 2007, 98(10), 1638-1642. [Content Brief]

[2]. Liu F, et al. Prolonged inhibition of class I PI3K promotes liver cancer stem cell expansion by augmenting SGK3/GSK-3β/β-catenin signalling. J Exp Clin Cancer Res. 2018 Jun 25;37(1):122. [Content Brief]

[3]. Wang P, et al. Class I PI3K inhibitor ZSTK474 mediates a shift in microglial/macrophage phenotype and inhibits inflammatory response in mice with cerebral ischemia/reperfusion injury. J Neuroinflammation. 2016 Aug 22;13(1):192. [Content Brief]

Help & FAQs

How should lyophilized recombinant proteins be reconstituted and stored?

1. Before opening the cap, centrifuge the vial at 13000 rpm for 20-30 seconds. This step will ensure that any lyophilized powder that may have adhered to the cap or walls is collected at the bottom of the vial, minimizing the risk of product loss.

Complete Stock Solution Preparation Table

Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.

Optional Solvent	Concentration Solvent Mass	1 mg	5 mg	10 mg	25 mg
DMSO	1 mM	2.3957 mL	11.9786 mL	23.9573 mL	59.8932 mL
	5 mM	0.4791 mL	2.3957 mL	4.7915 mL	11.9786 mL
	10 mM	0.2396 mL	1.1979 mL	2.3957 mL	5.9893 mL
	15 mM	0.1597 mL	0.7986 mL	1.5972 mL	3.9929 mL
	20 mM	0.1198 mL	0.5989 mL	1.1979 mL	2.9947 mL
	25 mM	0.0958 mL	0.4791 mL	0.9583 mL	2.3957 mL
	30 mM	0.0799 mL	0.3993 mL	0.7986 mL	1.9964 mL
	40 mM	0.0599 mL	0.2995 mL	0.5989 mL	1.4973 mL
	50 mM	0.0479 mL	0.2396 mL	0.4791 mL	1.1979 mL
	60 mM	0.0399 mL	0.1996 mL	0.3993 mL	0.9982 mL

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ZSTK474

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Publications Citing Use of MedChemExpress (MCE) ZSTK474

All PI3K Isoforms

Biological Activity

Chemical Information

Publications (18)

Solvent & Solubility

Protocol

Purity & Documentation

References

Help & FAQs

Complete Stock Solution Preparation Table

ZSTK474 Related Classifications

HY-50847 Related Classifications