CSF was obtained from KGC Research Institute (Daejeon, Korea). CSF was analyzed by HPLC and the composition of ginsenosides is as follows: 134 mg/g of G-Rg1, 106 mg/g of G-Re, 52 mg/g of G-Rf, 181 mg/g of G-Rb1, 165 mg/g of G-Rc, 108 mg/g of G-Rb2, 18 mg/g of Rb3, 72 mg/g of Rd, 44 mg/g of F2, 28 mg/g of Rg3. Collagen was obtained from Chrono-Log Co. (Havertown, PA, USA). Fura-2-acetoxymethyl ester (Fura-2/AM) and dimethyl sulfoxide (DMSO) were from Sigma Chemical Co. (St. Louis, MO, USA). Antibodies to phospho-p44/42 [p-extracellular signal-regulated protein kinase 1/2 (p-ERK1/2)], p44/42 (T-ERK1/2), phospho-p38 (p-p38), p38 (T-p38), phospho-SAPK (phospho-stress-activated protein kinase)/c-Jun N-terminal kinase (p-JNK), SAPK/JNK (T-JNK), phospho-phosphoinositide 3-kinase (phospho-PI3K; p-p85), PI3K (T-p38), phospho-mitogen-activated protein kinase kinase (p-MEK1/2), MEK1/2 (T-MEK1/2), phospho-Akt (p-Akt), and Akt (T-Akt) were acquired from Cell Signaling Technology (Beverly, MA, USA). Adenosine triphosphate (ATP) assay kits were purchased from Biomedical Research Service Center (Buffalo, NY, USA). Fibrinogen Alexa Fluor 488 conjugate was obtained from Molecular Probes (Eugene, OR, USA). All other chemicals were of reagent grade.
Fibrinogen alexa fluor 488 conjugate
Manufactured by Thermo Fisher Scientific
Sourced in United States
Fibrinogen Alexa Fluor 488 conjugate is a fluorescently labeled protein used for biological research applications. It is created by conjugating the Alexa Fluor 488 dye to the fibrinogen protein.
Automatically generated - may contain errors
Lab products found in correlation
Fura 2 am, by Merck Group (6 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (6 mentions)
Phospho sapk jnk, by Cell Signaling Technology (4 mentions)
Thrombin, by Chrono-Log (3 mentions)
Phospho p38, by Cell Signaling Technology (2 mentions)
Phospho akt, by Cell Signaling Technology (2 mentions)
Aspirin, by Merck Group (2 mentions)
Fibrinogen, by Merck Group (2 mentions)
Txb2 enzyme immunoassay eia kit, by Cayman Chemical (2 mentions)
β actin, by Cell Signaling Technology (2 mentions)
Eptifibatide, by Santa Cruz Biotechnology (2 mentions)
Anti β actin, by Santa Cruz Biotechnology (2 mentions)
Anti vasp, by Cell Signaling Technology (2 mentions)
Gr 144053, by Santa Cruz Biotechnology (2 mentions)
Lysis buffer, by Cell Signaling Technology (2 mentions)
Polyvinylidene difluoride pvdf membrane, by GE Healthcare (2 mentions)
Enhanced chemiluminesence solution ecl, by GE Healthcare (2 mentions)
Anti pi3k, by Cell Signaling Technology (2 mentions)
Fura 2 acetoxymethyl ester fura 2 am, by Merck Group (1 mentions)
κ carrageenan, by Merck Group (1 mentions)
10 protocols using fibrinogen alexa fluor 488 conjugate
1
Ginsenoside Composition and Signaling Analysis
2
Collagen-Mediated Platelet Activation
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Collagen was purchased from Chrono-log (Havertown, PA, USA). ASA, Fura-2/AM, κ-carrageenan, and dimethyl sulfoxide (DMSO) were obtained from Sigma-Aldrich (St. Louis, MO, USA). ATP Assay Kit was obtained from Biomedical Research Service Center (Buffalo, NY, USA). Fibrinogen Alexa Fluor 488 conjugate was obtained from Molecular Probes (Eugene, OR, USA). Antibodies against phospho-p44/42, p44/42, phospho-p38, p38, phospho-SAPK/JNK, SAPK/JNK, phospho-Akt, and Akt were acquired from Cell Signaling Technology (Beverly, MA, USA). HPLC-grade reagents, acetonitrile, and water were obtained from J. T. Baker (Phillipsburg, NJ, USA). All chemicals were of reagent-grade.
3
Platelet Signaling Pathway Activation
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Collagen, ADP, and thrombin were purchased from Chrono-log Corp. (Havertown, PA, USA). Fura-2/AM and dimethyl sulfoxide (DMSO) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Fibrinogen Alexa Fluor® 488 conjugate was purchased from Molecular Probes (Eugene, OR, USA), and the ATP assay kit was obtained from Biomedical Research Service Center (Buffalo, NY, USA). Antibodies against phospholipase Cγ2 (PLCγ2), phospho-p44/42 (phospho-extracellular signal-regulated kinase (ERK)), p44/42 (ERK), phospho-p38, p38, MEK, phospho-MEK, stress-activated protein kinase (SAPK)/c-Jun N-terminal protein kinase (JNK), phospho-SAPK/JNK, phospho-phosphoinositide 3-kinase (PI3K), PI3K, phospho-Akt, and Akt were acquired from Cell Signaling Technology (Beverly, MA, USA). Ultrapure water was obtained from J. T. Baker (Phillipsburg, NJ, USA). All chemicals were reagent grade.
4
Platelet Signaling Pathway Analysis
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Collagen was obtained from Chrono-Log Co. (Havertown, PA, USA). Aspirin, fibrinogen, plasmin and dimethyl sulfoxide (DMSO) were from Sigma Chemical Co. (St. Louis, MO, USA). Fura-2/AM was obtained from Sigma Chemical Co. (St. Louis, MO, USA). Antibodies against phospho-p44/42, p44/42, phospho-p38, p38, phospho-SAPK/JNK, phospho-PI3K (p85), phospho-Akt, and β-actin were acquired from Cell Signaling (Beverly, MA, USA). ATP assay kits were purchased from Biomedical Research Service Center (Buffalo, NY, USA). A TXB2 enzyme immunoassay (EIA) kit was from Cayman Chemical (Ann Arbor, MI, USA). fibrinogen Alexa Fluor 488 conjugate was obtained from Molecular Probes (Eugene, OR, USA). HPLC-grade reagents, methanol and water were obtained from J. T. Baker (Phillipsburg, NJ, USA). All other chemicals were of reagent grade.
5
Platelet Activation Pathway Analysis
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G-Ro was obtained from Ambo Institute (Daejon, Korea). Thrombin was obtained from Chrono-Log Corporation (Havertown, PA, USA). Anti-VASP, anti-phosphor-VASP (Ser157), anti-phosphor-VASP (Ser239), anti-rabbit IgG-HRP-horseradish peroxidase conjugate (HRP), and lysis buffer were purchased from Cell Signaling (Beverly, MA, USA). The αIIb/β3 inhibitor eptifibatide, GR 144053, and anti-β-actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Polyvinylidene difluoride membrane was purchased from GE Healthcare (Piseataway, NJ, USA). Enhanced chemiluminescence solution was purchased from GE Healthcare (Chalfont St. Giles, UK). cAMP and cyclic guanosine monophosphate (cGMP) enzyme immunoassay kits were purchased from Cayman Chemical (Ann Arbor, MI, USA). An A-kinase inhibitor Rp-8-Br-cAMPS, an A-kinase activator 8-(4-chlorophenylthio)-cAMP (pCPT-cAMP), and a G-kinase activator 8-Br-cGMP were purchased from Sigma Chemical Corporation (St. Louis, MO, USA). Fibrinogen Alexa Fluor 488 conjugate was obtained from Invitrogen Molecular Probes (Eugene, OR, USA).
6
Rg3-Enriched Red Ginseng Extract Characterization
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Rg3-RGE was obtained from Korea Ginseng Corporation (Daejeon, Korea). In brief, red ginseng (stem/root = 75:25) was extracted with water followed by extraction with 55% ethanol. A concentrated extract was prepared by multiple extractions. Consequently, the extract was subjected to GC-MS and the resulting extract constituent profile is shown in Table 1 , which confirms that the extract is Rg3-enriched red ginseng extract. Collagen was obtained from Chrono-Log Co. (Havertown, PA, USA). Fura-2/AM and dimethyl sulfoxide were purchased from Sigma Chemical Co. (St. Louis, MO, USA). ATP (adenosine triphosphate) assay kits were obtained from Biomedical Research Service Center (Buffalo, NY, USA). Fibrinogen Alexa Fluor 488 conjugate was purchased from Molecular Probes (Eugene, OR, USA). Antibodies directed against total-ERK1/2 (p44/42), phospho-ERK1/2 (p44/42), total-JNK, phospho-JNK, total-p38, phosphor-p38, total-PI3K (phosphatidylinositol 3 kinase) p85/p55, phospho-PI3K p85/p55, total-Akt (Ser473), phospho-Akt (Ser473), total-MKK4 (mitogen-activated protein kinase kinase 4), and phospho-MKK4 were purchased from Cell Signaling (Beverly, MA, USA). All chemicals used were of reagent grade.Table 1
Rg3-enriched red ginseng extract constituent profile
| Ginsenosides | Contents (mg/g) |
|---|---|
| Rb1 | 3.86 |
| 20(S)-Rg3 | 44.91 |
| Rc | 1.20 |
| Rb2 | 1.53 |
| Rd | 1.60 |
| Rf | 1.28 |
| Rh1 | 3.71 |
| 20(S)-Rg2 | 3.55 |
| 20(R)-Rg3 | 6.78 |
| Total | 67.41 |
7
Investigating Integrin-Mediated Cell Adhesion
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G-Ro (molecular weight=957.1) was obtained from Ambo Institute (Daejon, Korea). Thrombin was obtained from Chrono-Log Corporation (Havertown, PA, USA). Anti-PI3K, anti-phosphor-PI3K p85 (Tyr458), anti-Akt, anti-phosphor-Akt (Ser473), anti-rabbit IgG-horseradish peroxidase, and lysis buffer were purchased from Cell Signaling (Beverly, MA, USA). Wortmannin (PI3K inhibitor) and miltefosine (Akt inhibitor) were purchased from Cayman Chemical (Ann Arbor, MI, USA). CytoSelect 48-well cell adhesion assay kits (fibronectin-coated, colorimetric format) were purchased from Cell Biolabs (San Diego, CA, USA). Eptifibatide (αIIb/β3 inhibitor), GR 144053 (αIIb/β3 inhibitor), and anti-β-actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Polyvinylidene difluoride (PVDF) membrane and enhanced chemiluminesence solution (ECL) were purchased from General Electric Healthcare (Amersham, Buckinghamshire, UK). Fibrinogen Alexa Fluor 488 conjugate was obtained from Invitrogen Molecular Probes (Eugene, OR, USA).
8
Collagen-Mediated Platelet Activation Signaling
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Collagen was purchased from Chrono-Log Co. (Havertown, PA., USA). ATP assay kit was purchased from Biomedical Research Service Center (Buffalo, NY., USA). Cordycepin, A-kinase inhibitor Rp-8-Br-cAMPS, cGMP-dependent protein kinase (G-kinase) inhibitor Rp-8-Br-cGMPS, A-kinase activator pCPT-cAMP, and G-kinase activator 8-Br-cGMP were obtained from Sigma Chemical Corporation (St. Louis, MO., USA). Serotonin ELISA kit was purchased from Labor Diagnostika Nord GmbH & CO. (Nordhorn, Germany). Anti-VASP, anti-phosphor- VASP (Ser157), anti-phosphor-VASP (Ser239), anti-PI3K, anti-phosphor- PI3K, anti-Akt, anti-phosphor-Akt, and anti-rabbit IgG-horseradish peroxidase conjugate (HRP), and lysis buffer were obtained from Cell Signaling (Beverly, MA., USA). Poly-vinylidene difluoride (PVDF) membrane was from GE Healthcare (Piseataway, NJ., USA). Enhanced chemiluminesence solution (ECL) was from GE Healthcare (Chalfont St., Giles, Buckinghamshire, UK). Fibrinogen Alexa Fluor 488 conjugate was obtained from Invitrogen Molecular Probes (Eugene, OR., USA).
9
Collagen-Induced Platelet Activation
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Collagen was obtained from Chrono-Log Co. (Havertown, PA, USA). Aspirin, fibrinogen, dimethyl sulfoxide (DMSO), and Fura-2/AM were obtained from Sigma Chemical Co. (St. Louis, MO, USA). Antibodies of phospho-p38, p38, phospho-SAPK/JNK, phospho-PI3 K (p85), class I PI3 K and isoforms, and β-actin were purchased from Cell Signaling (Beverly, MA, USA). ATP (adenosine triphosphate) assay kit was purchased from Biomedical Research Service Center (Buffalo, NY, USA). TXB2 enzyme immunoassay (EIA) kit was purchased from Cayman Chemical (AnnArbor, MI, USA). fibrinogen Alexa Fluor 488 conjugate was purchased from Molecular Probes (Eugene, OR, USA).
10
Platelet Activation Signaling Pathways
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Thrombin and Collagen (Native collagen fibrils, type I, from equine tendons) were procured from Chrono-Log Co. (Havertown, PA, USA). ATP assay kit was acquired from Biomedical Research Service Center (Buffalo, NY, USA), while Fibrinogen Alexa Fluor® 488 conjugate was acquired from Molecular Probes (Eugene, OR, USA). Fura-2/AM and dimethyl sulfoxide (DMSO) were obtained from Sigma Chemical Co. (St. Louis, MO, USA). Antibodies against ERK (p44/42), phospho-ERK (p44/42), JNK, phospho-JNK, PI3K (p85/p55), phospho-PI3K (p85/p55), Akt (Ser473), phospho-Akt (Ser473), MKK4, phosphor-MKK4, Src (Tyr416), and phospho-Src (Tyr416) were obtained from Cell signaling (Beverly, MA, USA). Water was acquired from J. T. Baker (Phillipsburg, NJ, USA). All chemicals were of reagent grade.
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