Log In Sign up
The largest database of trusted experimental protocols

Histopaque 1077 medium

Manufactured by Merck Group
Visit Supplier
Sourced in United States

Histopaque 1077 is a density gradient medium used for the isolation of mononuclear cells from blood. It is a sterile, endotoxin-tested solution consisting of a polysucrose and sodium diatrizoate, adjusted to a density of 1.077 g/mL.

Automatically generated - may contain errors

Lab products found in correlation

L glutamine, by GE Healthcare (2 mentions) Penicillin, by Corning (2 mentions) Streptomycin, by Corning (2 mentions) M mlv reverse transcriptase, by Thermo Fisher Scientific (2 mentions) Digoxin, by Merck Group (1 mentions) Resazurin, by Merck Group (1 mentions) Lc ms grade acetonitrile, by Merck Group (1 mentions) Methanol isocratic grade, by Merck Group (1 mentions) Milli q water purification system, by Merck Group (1 mentions) 2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (1 mentions) Thiobarbituric acid, by Merck Group (1 mentions) Indomethacin, by Merck Group (1 mentions) Penicillin streptomycin solution, by Merck Group (1 mentions) Lc ms grade formic acid, by Merck Group (1 mentions) Chloroform, by Merck Group (1 mentions) Phosphate buffered saline (pbs), by Merck Group (1 mentions) Cfx384, by Bio-Rad (1 mentions) Hot firepol evagreen qpcr mix plus, by Solis BioDyne (1 mentions) Rneasy mini kit, by Qiagen (1 mentions)

Close spelling variants of this product

Histopaque®-1077 density gradient cell separation medium Histopaque (Histopaque-1077 Cell Separation Medium, Histopaque-1077 cell separation medium Histopaque-1077 Histopaque medium Histopaque

7 protocols using histopaque 1077 medium

1

Peroxynitrite Synthesis and Antioxidant Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols
Chloroform, methanol (isocratic grade) and acetonitrile (LC–MS grade) were purchased from Merck (Darmstadt, Germany). Formic acid (LC–MS grade) and digoxin (internal standard, IS) were purchased from Sigma–Aldrich (St. Louis, MO, USA). Ultrapure water was prepared using a Milli–Q water purification system (Millipore, Milford, CT, USA). Peroxynitrite was synthesized according to the method of Pryor et al. [40 (link)]. DPPH, Trolox®, rosmarinic acid, thiobarbituric acid, Histopaque®–1077 medium, indomethacin, penicillin–streptomycin solution for cell culture and resazurin (In vitro toxicology assay kit, resazurin based, TOX8–1KT) were also purchased from Sigma–Aldrich (St. Louis, MO, USA). COX (human) Inhibitor Screening Assay Kit (Item No. 701230) and COX Colorimetric Inhibitor Screening Assay Kit (Item No. 701050) were from Cayman Chemicals (Ann Arbor, MI, USA). Reagents and cuvettes for cytotoxicity assays were purchased from NanoEnTek Inc. (Seoul, Korea). All other reagents were provided by local or international suppliers (mainly Avantor Performance Materials, Gliwice, Poland).
Krzyżanowska-Kowalczyk J., Kowalczyk M., Ponczek M.B., Pecio Ł., Nowak P, & Kolodziejczyk-Czepas J. (2021). Pulmonaria obscura and Pulmonaria officinalis Extracts as Mitigators of Peroxynitrite-Induced Oxidative Stress and Cyclooxygenase-2 Inhibitors–In Vitro and In Silico Studies. Molecules, 26(3), 631.
+ Open protocol
+ Expand
2

Hematopoietic Cell Lines and Patient Samples for AML/CML Study

Check if the same lab product or an alternative is used in the 5 most similar protocols
We used twelve human malignant hematopoietic cell lines of various lineages (ATCC), including seven myeloid (HEL, K-562, THP-1, U937, KG-1a, HL-60, DAMI) and five lymphoid (DAUDI, RAJI, NALM-6, JURKAT, MOLT4) cell lines. All cell lines were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium containing L-glutamine (GE Healthcare), 10% heat-inactivated fetal bovine serum (FBS; Seradigm), 100 units/mL penicillin, and 10 μg/mL streptomycin (Corning) and cultured in a humidified atmosphere of 5% CO2 at 37°C, with an exchange of medium every 48 h.
Ten patients with recently diagnosed AML (n=6) and CML (n=4) were obtained for this study according to Institutional IRB guidelines. EDTA-anticoagulated whole blood was obtained from these patients, and peripheral blood mononuclear cells (PB-MNCs) were immediately separated by density-gradient centrifugation using Histopaque 1077 medium (Sigma-Aldrich).
Abdelbaset-Ismail A., Borkowska-Rzeszotek S., Kubis E., Bujko K., Brzeźniakiewicz-Janus K., Bolkun L., Kloczko J., Moniuszko M., Basak G.W., Wiktor-Jedrzejczak W, & Ratajczak M.Z. (2016). Activation of the complement cascade enhances motility of leukemic cells by downregulating expression of HO-1. Leukemia, 31(2), 446-458.
+ Open protocol
+ Expand
3

Isolation of Peripheral Blood Lymphocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols
In an Eppendorf tube, 20 μL of whole peripheral blood was mixed with 1 mL of RPMI 1640 culture medium and then layered on 200 μL of Histopaque-1077 medium (Sigma-Aldrich, Poznan, Poland). The samples were centrifuged for 3 min at 500× g. The separated fraction containing lymphocytes and monocytes was transferred to freshly prepared 1 mL RPMI 1640 medium. The sample was centrifuged again for 3 min at 500× g. At the end of the isolation, the lymphocyte pellet was resuspended in 100 μL of 1% PBS (potassium buffer solution) (Sigma-Aldrich, Poznan, Poland).
Krystyjan M., Khachatryan G., Khachatryan K., Konieczna-Molenda A., Grzesiakowska A., Kuchta-Gładysz M., Kawecka A., Grzebieniarz W, & Nowak N. (2022). The Functional and Application Possibilities of Starch/Chitosan Polymer Composites Modified by Graphene Oxide. International Journal of Molecular Sciences, 23(11), 5956.
+ Open protocol
+ Expand
4

Isolating PB-MNCs from Newly Diagnosed AML Patients

Check if the same lab product or an alternative is used in the 5 most similar protocols
Patients with newly diagnosed AML (n=7) were employed in this study. Such
a diagnosis was made according to the WHO classification system [40 (link)]. Determination of complete blood counts and flow
cytometry were performed to confirm the existence of blast cells.
EDTA-anticoagulated whole blood was collected from these AML patients, and the
peripheral blood mononuclear cells (PB-MNCs) were immediately separated by
density-gradient centrifugation using Histopaque 1077 medium (Sigma-Aldrich).
Patients samples were obtained for this study according to Institutional IRB
guidelines.
Abdelbaset-Ismail A., Cymer M., Borkowska-Rzeszotek S., Brzeźniakiewicz-Janus K., Rameshwar P., Kakar S.S., Ratajczak J, & Ratajczak M.Z. (2018). Bioactive Phospholipids Enhance Migration and Adhesion of Human Leukemic Cells by Inhibiting Heme Oxygenase 1 (HO-1) and Inducible Nitric Oxygenase Synthase (iNOS) in a p38 MAPK-Dependent Manner. Stem Cell Reviews, 15(1), 139-154.
+ Open protocol
+ Expand
5

PBMC Isolation and qPCR Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Peripheral blood mononuclear cells (PBMCs) were isolated by density gradient centrifugation using Histopaque 1077 medium (Sigma Aldrich, St. Louis, MO, USA), washed twice with PBS containing 2 mM EDTA and snap frozen. Samples were collected at both locations but transferred from Helsinki to Tampere on dry ice for further preparations and analysis. Total RNA was extracted from PBMCs using the RNeasy Mini-Kit (Qiagen, Valencia, CA, USA). Total RNA (0.5 µg) was reverse-transcribed using M-MLV reverse transcriptase (Thermo Scientific, Waltham, MA, USA) according to the manufacturer’s instructions. Quantitative PCR (qPCR) reactions were performed by using HOT FIREPol EvaGreen qPCR Mix Plus (Solis BioDyne, Tartu, Estonia). Primer sequences for STAT1, STAT3, STAT4, STAT5A, STAT5B, STAT6, JAK1, JAK2, JAK3, TYK2, SOCS1, SOCS2, SOCS3, CIS1 and β-actin are listed in Supplementary Table 3. The 10-µl real-time PCR reactions were performed with CFX384 (Bio-Rad Laboratories, Hercules, CA, USA) and gene expression was quantified by using the delta C(T) method by normalizing to the expression of β-actin.
Palmroth M., Kuuliala K., Peltomaa R., Virtanen A., Kuuliala A., Kurttila A., Kinnunen A., Leirisalo-Repo M., Silvennoinen O, & Isomäki P. (2021). Tofacitinib Suppresses Several JAK-STAT Pathways in Rheumatoid Arthritis In Vivo and Baseline Signaling Profile Associates With Treatment Response. Frontiers in Immunology, 12, 738481.
+ Open protocol
+ Expand
6

Isolation of AML Patient PBMCs

Check if the same lab product or an alternative is used in the 5 most similar protocols
Ten patients with newly diagnosed acute myeloid leukemia (AML) were recruited for the study. A diagnosis of AML was established based on the WHO classification system [31 (link)]. Complete blood counts and flow cytometry were performed in order to confirm the presence of blast cells. Detailed clinical, phenotypical, and molecular characteristic of recruited AML patients are presented in supplemental Table 1. EDTA-anticoagulated whole blood was obtained from these AML patients and immediately processed. Peripheral blood mononuclear cells (PB- MNCs) were isolated by means of density-gradient centrifugation using Histopaque 1077 medium (Sigma-Aldrich).
Abdelbaset-Ismail A., Borkowska S., Janowska-Wieczorek A., Tonn T., Rodriguez C., Moniuszko M., Bolkun L., Koloczko J., Eljaszewicz A., Ratajczak J., Ratajczak M.Z, & Kucia M. (2015). Novel evidence that pituitary gonadotropins directly stimulate human leukemic cells-studies of myeloid cell lines and primary patient AML and CML cells. Oncotarget, 7(3), 3033-3046.
+ Open protocol
+ Expand
7

Malignant Hematopoietic Cell Lines and Primary AML/CML Samples

Check if the same lab product or an alternative is used in the 5 most similar protocols
We used twelve human malignant hematopoietic cell lines of various lineages (ATCC), including seven myeloid (HEL, K-562, THP-1, U937, KG-1a, HL-60, DAMI) and five lymphoid (DAUDI, RAJI, NALM-6, JURKAT, MOLT4) cell lines. All cell lines were cultured in Roswell Park Memorial Institute (RPMI) 1640 maedium containing l-glutamine (GE Healthcare, South Logan, UT, USA), 10% heat-inactivated fetal bovine serum (FBS; Seradigm, Radnor, PA, USA), 100 units/ml penicillin and 10 μg/ml streptomycin (Corning Costar, Corning, NY, USA), and cultured in a humidified atmosphere of 5% CO2 at 37 °C, with an exchange of medium every 48 h.
Ten patients with recently diagnosed acute myeloid leukemia (AML) (n=6) and chronic myeloid leukemia (CML) (n=4) were obtained for this study according to Institutional IRB guidelines. Ethylenediaminetetraacetic acid-anticoagulated whole blood was obtained from these patients, and peripheral blood mononuclear cells (PB-MNCs) were immediately separated by density-gradient centrifugation using Histopaque 1077 medium (Sigma-Aldrich, St Louis, MO, USA).
Abdelbaset-Ismail A., Borkowska-Rzeszotek S., Kubis E., Bujko K., Brzeźniakiewicz-Janus K., Bolkun L., Kloczko J., Moniuszko M., Basak G.W., Wiktor-Jedrzejczak W, & Ratajczak M.Z. (2016). Activation of the complement cascade enhances motility of leukemic cells by downregulating expression of HO-1. Leukemia, 31(2), 446-458.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!