LPA, EGF, IGF and PTx were from Sigma. Human SDF1-α was from Pepro Tech. Growth-factor-reduced Matrigel was from BD Biosciences. Rabbit anti-AKT, mouse anti-phospho-AKT, rabbit anti-ERK1/2, and mouse anti-phospho-ERK1/2 antibodies were from Cell Signaling Technology. Mouse anti-mortalin was from NeuroMab. Mouse anti-Gαt was a gift from Dr. Heidi Hamm (Vanderbilt University). Human allophycocyanin-conjugated CD44 was from BD Biosciences, and human phycoerythrin-conjugated CD133 was from Miltenyl Biotech. Paclitaxel was from LC Laborateries. Gallein was from TCI America.
Human sdf 1α
Manufactured by Thermo Fisher Scientific
Sourced in United States
Human SDF-1α is a recombinant protein that represents the chemokine Stromal Cell-Derived Factor 1 alpha. It is a small cytokine that plays a role in the regulation of cell migration and adhesion.
Automatically generated - may contain errors
Lab products found in correlation
Reverse transcription kit, by Thermo Fisher Scientific (3 mentions)
Nigericin, by Merck Group (2 mentions)
Wortmannin, by Apexbio (2 mentions)
Gallein, by Bio-Techne (2 mentions)
Swainsonine, by Merck Group (2 mentions)
Rapamycin, by Merck Group (2 mentions)
Monensin, by Merck Group (2 mentions)
Amd3100, by Santa Cruz Biotechnology (2 mentions)
Uk14304, by Thermo Fisher Scientific (2 mentions)
As 604850, by Apexbio (2 mentions)
Gsk2292767, by Apexbio (2 mentions)
Tgx 221, by Adooq (2 mentions)
Hs 173, by Adooq (2 mentions)
Transwell chamber, by Corning (2 mentions)
Mouse anti phospho erk1 2, by Cell Signaling Technology (1 mentions)
Rabbit anti akt, by Cell Signaling Technology (1 mentions)
Rabbit anti erk1 2, by Cell Signaling Technology (1 mentions)
Human allophycocyanin conjugated cd44, by BD (1 mentions)
Epidermal growth factor (egf), by Merck Group (1 mentions)
Mouse anti phospho akt, by Cell Signaling Technology (1 mentions)
8 protocols using human sdf 1α
1
Signaling Pathways in Cancer Stem Cells
2
Investigating SDF1α-Mediated Cell Migration
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Human SDF1α was purchased from PeproTech; UK14304, rapamycin, brefeldin A, monensin, nigericin, swainsonine, and Exo2 were from Sigma–Aldrich; pertussis toxin was from List Biological Laboratories; gallein was from Tocris Bioscience; wortmannin, AS-604850, and GSK2292767 were from ApexBio; TGX-221 and HS-173 were from Adooq Bioscience; secinH3, GCA, nocodazole, ilimaquinone, AMD3100, control siRNA, and siRNA targeting human ARF1, CRISPR–Cas9 control plasmids, and knockout plasmids targeting human ARF1, and antibodies against phospho-ERK1/2 and β-actin were from Santa Cruz Biotechnology; p230 antibodies were from BD Transduction Laboratories; ARF1 antibodies were purchased from Abcam; ERK1/2 antibodies were from Cell Signaling Technology; 12-well inserts and Matrigel matrix were from Corning. All other materials were obtained as described elsewhere (9 (link), 19 (link), 27 (link)).
3
Bioactive Scaffold Functionalization
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Sterilized electrospun PEGdma-PLA scaffolds were either coated with 0.6 µg/mL human SDF-1α (300-28 A, Peprotech, Rocky Hill, US) or 100 µg/mL of the generated human DCN. To enable protein adsorption, scaffolds were incubated for 4 hours at 37 °C with the protein solutions. Unbound protein was removed by washing the functionalized scaffolds with sterile 1x PBS.
4
Glioma Cell Lines and Peptide Treatments
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Human glioma cell lines SHG-44 and U251 were purchased from Cell Bank of the Chinese Academy of Sciences (Shanghai, China). NT21MP was designed by our laboratory and synthesized by GL Biochem Ltd. (Shanghai, China). The amino acid sequence information of the NT21MP is H-D-leu-D-Gly-D-Ala-D-Ser-D-Trp-D-His-D-Arg-D-Pro-D-Asp-D-Lys-Cys-Cys-Leu-Gly-Tyr-Gln-Lys-Arg-Pro-Leu-Pro-OH. Human-SDF-1α was purchased from PeproTech (Rocky Hill, NJ, USA). AMD3100 and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Primary antibodies against Bcl-2, Bax, caspase-3, cyclin D1 and β-actin were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). A mouse anti-human CXCR4 mAb was purchased from Abcam (clone: 44716.111). Secondary antibodies conjugated to horseradish peroxidase (HRP) were purchased from ZSGB-Bio, Co., Ltd. (Beijing, China). Apoptosis kit was obtained from BD Biosciences (San Jose, CA, USA). Hoechst 33258 was purchased from Sigma-Aldrich. Reverse transcription kit was obtained from Thermo Fisher Scientific (Waltham, MA, USA) and the SYBR Premix Dimer Eraser™ reagent kit from Takara, Co., Ltd. (Shiga, Japan).
5
Quantifying Chemotaxis Towards SDF-1α
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The chemotaxis assay was performed using a transwell chamber (5 µm; Corning). 200,000 cells suspended in 100 µl medium were placed into the top chamber, and 600 µl medium containing 50 ng/ml human SDF-1α (PeproTech) was added to the bottom well. After 4 h of incubation, cells in the bottom well were collected and the cell number was counted using an Accuri C6 (BD).
6
Memantine Modulates T Cell Migration
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Isolated CD3+ cells (4x106) remained untreated or were pre-incubated with memantine (20 and 50 μM) for 30 min in AIM-V medium (Gibco®, Life Technologies) supplemented with 0.1% bovine serum albumin and 10 mM HEPES (pH 7.4) and then transferred unto uncoated or fibronectin-coated (6.5 μg/ml; Roche Diagnostics, Basel, Switzerland) transwell chambers (6.5 mm diameter and 3.0 μm pore; Corning Costar, Tewksbury, MA). Cells were allowed to migrate towards human SDF-1α (100 ng/ml; PeproTech, Hamburg, Germany) for 150 min at 37°C. Migration in the absence of chemokine served as a control. Migration was stopped by the addition of 0.1 M EDTA. Migrated cells were stained with CD4-PE and CD8-FITC Abs (BD Pharmingen, Heidelberg, Germany) and acquired for 30 sec at a FACSFortessaTM (BD Bioscience, Heidelberg, Germany). Relative migration was determined by defining the number of cells that migrated in the absence of memantine (mem. 0 μM) as 100%.
7
Investigating PI3Kγ Signaling Pathways
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Human SDF1α was purchased from PeproTech; UK14304, GW5074, rapamycin, BFA, ilimaquinone, monensin, nigericin, swainsonine, and LY294002 were from Sigma Aldrich; nocodazole, insulin-like growth factor 1, AMD3100, control siRNA (medium GC), siRNAs targeting to human PI3Kγ regulatory subunits p101 and p87, and antibodies against GFP, phospho-ERK1/2, Gγ9, and β-actin and the PI3Kγ subunits p110γ, p101 and p87 were from Santa Cruz Biotechnology; wortmannin, AS-604850, and GSK2292767 were from ApexBio; TGX-221 and HS-173 were from Adooq Bioscience; U0126 and PD98059 were from Calbiochem; EGF, puromycin, and blasticidin S were from Thermo Fisher Scientific; U-73122 and AZD5363 were from MedChemExpress; U-73433, CRT0066101, and Go6976 were from Cayman Chemical; PTX was from List Biological Laboratories; gallein and rauwolscine were from Tocris Bioscience; D-Luciferin was from GoldBio; antibodies against hemagglutinin and ERK1/2 were from Cell Signaling Technology; antibodies against Gγ3 were from Abcam. All other materials were obtained as described (52 (link), 53 (link)).
8
Transwell B-cell Migration Assay
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Human B-cells were selected from murine spleens using CD20+ microbead kit (Miltenyi Biotec) according to the manufacturer’s instructions. Purified B-cells were cultured overnight at 37 °C/5% CO2 in complete RPMI medium (FCS [10%], Penicillin-Streptomycin-Glutamine [1×]). In the next day, 50,000 viable cells were re-suspended in 100 µl of complete RPMI and seeded on 5 µM pore size Transwell inserts (Sigma-Aldrich). The lower chamber of the 24-well plate was filled with 600 µl of complete RPMI medium supplemented with human SDF-1α (250 ng/ml, Peprotech). After 37 °C/5% CO2 incubation for 3 h, the transmigrated cells collected in the lower chamber were stained with anti-human CD19 PerCp Cy5.5 antibody and analysed by FACS. The percentage of migration was estimated as (total number of viable CD19+ cell in the lower chamber)/(total number of initial cell in the upper chamber) × 100.
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