Log In Sign up
The largest database of trusted experimental protocols

Stem cell factor (scf)

Manufactured by BioLegend
Visit Supplier
Sourced in United States, United Kingdom, Canada

SCF is a recombinant protein that functions as a cytokine, promoting the proliferation and differentiation of hematopoietic stem cells.

Automatically generated - may contain errors

Lab products found in correlation

Interleukin 3 (il 3), by BioLegend (12 mentions) Interleukin 6 (il 6), by BioLegend (7 mentions) Gm csf, by BioLegend (6 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (6 mentions) 37 c incubator, by Thermo Fisher Scientific (6 mentions) Iscove modified dulbecco media (imdm), by Thermo Fisher Scientific (5 mentions) Flt3 ligand, by BioLegend (4 mentions) G csf, by BioLegend (3 mentions) Interleukin 7 (il 7), by BioLegend (3 mentions) Glutamax, by Thermo Fisher Scientific (3 mentions) 2 mercaptoethanol, by Merck Group (3 mentions) Recombinant mouse il 3, by BioLegend (3 mentions) Facsymphony a5, by BD (3 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions) Mycoalert plus kit, by Lonza (2 mentions) Es cell grade fbs, by Merck Group (2 mentions) Moflo xdp, by Beckman Coulter (2 mentions) Alpha mem, by Thermo Fisher Scientific (2 mentions) Gm csf, by R&D Systems (2 mentions)

36 protocols using stem cell factor (scf)

1

Culturing Primary Cells and CML Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols
Primary cells were cultured in SFM supplemented with stem cell factor (SCF; 0.2 ng/ml; BioLegend, #573902), granulocyte colony-stimulating factor (G-CSF; 1 ng/ml; BioLegend, #578602), granulocyte-macrophage CSF (GM-CSF; 0.2 ng/ml; BioLegend, #572902), interleukin-6 (IL6; 1 ng/ml; BioLegend, #570802), macrophage inflammatory protein α (MIPα; 0.2 ng/ml; PeproTech, #300-08), leukemia inhibitory factor (LIF; 0.05 ng/ml; PeproTech, #300-05), [bovine serum albumin (BSA), insulin, and transferrin (BIT)] (20%; STEMCELL Technologies, #09500), low-density lipoprotein (40 μg/ml; Sigma-Aldrich, #L4646), 2-mercaptoethanol (0.1 mM; Invitrogen, #31350-010), and 1% penicillin/streptomycin (LifeTech, #15140-122) and resuspended in Iscove’s modified Dulbecco’s medium (LifeTech, #12440-053). The CML cell lines K562 and KCL22 (DSMZ) were cultured in RPMI 1640 medium (LifeTech, #11875-093) supplemented with 1% penicillin/streptomycin, 1% l-glutamine, and 10% (v/v) fetal calf serum. Both primary cells and cell lines were passaged every 2 to 3 days, maintained at a concentration of 2 × 105 cells/ml at 37°C with 5% CO2, and routinely tested for mycoplasma. ULK1 KO single-cell cloning was performed with serial dilution in a 96-well plate (100 μl per well). Puromycin selection (3 μg/ml) was performed after cell expansion, and Western blotting was performed to validate ULK1 KO.
Ianniciello A., Zarou M.M., Rattigan K.M., Scott M., Dawson A., Dunn K., Brabcova Z., Kalkman E.R., Nixon C., Michie A.M., Copland M., Vetrie D., Ambler M., Saxty B, & Helgason G.V. (2021). ULK1 inhibition promotes oxidative stress–induced differentiation and sensitizes leukemic stem cells to targeted therapy. Science translational medicine, 13(613), eabd5016.
+ Open protocol
+ Expand
2

Differentiation potential of single MPP cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
To analyze the differentiation potential of MMPs, single MPP cells were sorted by MoFlo XDP (Beckman Coulter) and plated (one cell/well) in the MEM alpha (Life technology) medium containing 20% ES cell-grade FBS (Millipore), SCF (50 ng/ml: BioLegend), and Flt3 ligand (30 ng/ml: BioLegend) on the layer of OP9 cells in 96-well plates as previously described33 (link). The OP9 cell line was obtained from ATCC and confirmed to be free of mycoplasma by using the MycoAlert PLUS kit (Lonza). Two days after cell plating, IL-3 (10 ng/ml; BioLegend), IL-7 (10 ng/ml; BioLegend), and GM-CSF (10 ng/ml; R&D Systems) were added to each well to induce cell differentiation. “Plating efficiency” denotes cells successfully expanded in culture, determined by microscopic observation. Expanded cells were analyzed by flow cytometry to determine whether they developed into either granulocyte/macrophages (GM) (Gr-1+CD11b+B220−CD19− and Gr-1−CD11b+B220−CD19−) or B cells (Gr-1−CD11b−B220+CD19+).
Kanayama M., Xu S., Danzaki K., Gibson J.R., Inoue M., Gregory S.G, & Shinohara M.L. (2017). Skewing of the population balance of lymphoid and myeloid cells by secreted and intracellular osteopontin. Nature immunology, 18(9), 973-984.
+ Open protocol
+ Expand
3

Differentiation potential of single MPP cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
To analyze the differentiation potential of MMPs, single MPP cells were sorted by MoFlo XDP (Beckman Coulter) and plated (one cell/well) in the MEM alpha (Life technology) medium containing 20% ES cell-grade FBS (Millipore), SCF (50 ng/ml: BioLegend), and Flt3 ligand (30 ng/ml: BioLegend) on the layer of OP9 cells in 96-well plates as previously described33 (link). The OP9 cell line was obtained from ATCC and confirmed to be free of mycoplasma by using the MycoAlert PLUS kit (Lonza). Two days after cell plating, IL-3 (10 ng/ml; BioLegend), IL-7 (10 ng/ml; BioLegend), and GM-CSF (10 ng/ml; R&D Systems) were added to each well to induce cell differentiation. “Plating efficiency” denotes cells successfully expanded in culture, determined by microscopic observation. Expanded cells were analyzed by flow cytometry to determine whether they developed into either granulocyte/macrophages (GM) (Gr-1+CD11b+B220−CD19− and Gr-1−CD11b+B220−CD19−) or B cells (Gr-1−CD11b−B220+CD19+).
Kanayama M., Xu S., Danzaki K., Gibson J.R., Inoue M., Gregory S.G, & Shinohara M.L. (2017). Skewing of the population balance of lymphoid and myeloid cells by secreted and intracellular osteopontin. Nature immunology, 18(9), 973-984.
+ Open protocol
+ Expand
4

Profiling Cytokine Secretion in Breast Cancer

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Different cytokines and growth factors known to either chemoattract or activate MCs were measured in the conditioned media of the BrC lines. Levels of Regulated on Activation, Normal T Cell Expressed and Secreted (RANTES/CCL5), stromal cell-derived factor 1 (SDF1/CXCL12), granulocyte-colony-stimulating factor (G-CSF), granulocyte macrophage-colony-stimulating factor (GM-CSF) and monocyte chemoattractant protein-1 (MCP1/CCL2) were measured using the MILLIPLEX HCYTOMAG-60K kit (EMD Millipore, Darmstadt, Germany) [32 (link)]. Levels of IL-8 (BD, San Diego, CA, USA) and SCF (Biolegend, San Diego, CA, USA) were determined through Enzyme-Linked ImmunoSorbent Assays (ELISA) and following the manufacturer´s recommended procedure.
Aponte-López A., Enciso J., Muñoz-Cruz S, & Fuentes-Pananá E.M. (2020). An In Vitro Model of Mast Cell Recruitment and Activation by Breast Cancer Cells Supports Anti-Tumoral Responses. International Journal of Molecular Sciences, 21(15), 5293.
+ Open protocol
+ Expand
5

Transduction of Roryd-/- Bone Marrow

Check if the same lab product or an alternative is used in the 5 most similar protocols
Retrovirus for transducing
Rroid−/− bone marrow was
generated as previously described (Kurachi
et al., 2014). Briefly, Id2 cDNA was cloned into
the MSCV-IRES-GFP plasmid. Empty or Id2-expressing retrovirus were produced
in 293T human embryonic kidney cells with MSCV expression and pCL-Eco
packaging plasmids using Lipofectamine 3000 (Invitrogen) according to the
manufacturer’s instructions.
Bone marrow for transduction was isolated by flushing femurs and
tibias from Rroid−/− mice 4 days
after treatment with 5mg 5-Fluorouracil. Bone marrow cells were seeded at
2.5×106 cells/mL in a 6-well plate in IMDM+
15% FBS+ 1X Penicillin/Streptomycin+ 10ng/mL
IL-3+ 5ng/mL IL-6+ and 100ng/mL SCF (Biolegend) and cultured
overnight. Cultures were readjusted to 5×106 cells/mL in
1mL culture medium, and 1mL of retroviral supernatants supplemented with
10μg/mL Prolybrene (EMD Millipore) was added to the appropriate
wells. Cells were spinfected at 230xg for 2h at room temperature, then
incubated at 37°C overnight. Transduced bone marrow was then washed
and transferred to lethally irradiated hosts. The day before transfer,
CD45.1+ congenic recipients were irradiated with
2×550rad given 3h apart. Mice were analyzed 6 weeks following
transfer.
Mowel W.K., McCright S.J., Kotzin J.J., Collet M.A., Uyar A., Chen X., DeLaney A., Spencer S.P., Virtue A.T., Yang E., Villarino A., Kurachi M., Dunagin M.C., Pritchard G.H., Stein J., Hughes C., Fonseca-Pereira D., Veiga-Fernandes H., Raj A., Kambayashi T., Brodsky I.E., O’Shea J.J., Wherry E.J., Goff L.A., Rinn J.L., Williams A., Flavell R.A, & Henao-Mejia J. (2017). Group 1 Innate lymphoid cell lineage identity is determined by a cis-regulatory element marked by a long non-coding RNA. Immunity, 47(3), 435-449.e8.
+ Open protocol
+ Expand
6

Isolation and Culture of Bone Marrow-Derived Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
For bone marrow-derived MC (BMMC) culture, WT bone marrow was obtained from WT C57BF/6 mice in ice-cold Hank's Balanced Salt Solution (HBSS, Gibco), and cultured in complete RPMI containing 10% fetal bovine serum (FBS) (Hyclone), 1 mM nonessential amino acids, 25 mM HEPES, 2 mM L-glutamine, 1 mM sodium pyruvate, 1× Antibiotic-Antimycotic (all reagents from Gibco), 10 ng/ml SCF and 5 ng/ml IL-3 (BioLegend) for 8 to 12 weeks. For BMDC culture, bone marrow was obtained from WT C57BL/6 or CD301b-DTR-GFP mice and cultured in RPMI media containing 10% FBS, 1× GlutaMAX® (Gibco), 20 ng/ml granulocyte macrophage colony-stimulating factor (GM-CSF) (BioLegend), or 20 ng/ml GM-CSF and 50 ng/ml IL-4 for 6-7 days. For bone marrow-derived macrophage (BMM) culture, the same procedures were followed replacing the growth factors with 20 ng/ml macrophage colony-stimulating factor (M-CSF). The rat MC RBL-2H3 cells (ATCC) were cultured in minimum essential medium (MEM) medium (Gibco) containing 15% FBS and antibiotics. JAWSII cells (ATCC) were maintained in MEM α medium (Gibco) supplemented with 20% FBS, 1 mM sodium pyruvate, 100 U/ml penicillin, and 100 μg/ml streptomycin. All cells were cultured at 37 °C in a humidified water-jacketed incubator under 5% CO2 / 95% air atmosphere.
Choi H.W., Suwanpradid J., Kim I.H., Staats H.F., Haniffa M., MacLeod A.S, & Abraham S.N. (2018). Perivascular Dendritic Cells Elicit Anaphylaxis by Relaying Allergens to Mast Cells via Microvesicles. Science (New York, N.Y.), 362(6415), eaao0666.
+ Open protocol
+ Expand
7

Comprehensive Hematopoietic Stem Cell Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols
Bulk or single cells were sorted into methylcellulose culture medium (Stemcell Technology). Myeloid colony-forming capacity was assessed by MethoCult™ GF M3534 for myeloid committed progenitors. HSC colony-forming capacity was tested by using MethoCult™ GF M3434. HSPC Pre-B colony formation was modified by supplementing MethoCult™ M3630 medium with recombinant mouse IL-7 (BioLegend) and SCF (BioLegend) as described previously.
Chen X., Deng H., Churchill M.J., Luchsinger L.L., Du X., Chu T.H., Friedman R.A., Middelhoff M., Ding H., Tailor Y.H., Wang A.L., Liu H., Niu Z., Wang H., Jiang Z., Renders S., Ho S.H., Shah S.V., Tishchenko P., Chang W., Swayne T.C., Munteanu L., Califano A., Takahashi R., Nagar K.K., Renz B.W., Worthley D.L., Westphalen C.B., Hayakawa Y., Asfaha S., Borot F., Lin C.S., Snoeck H.W., Mukherjee S, & Wang T.C. (2017). Bone marrow myeloid cells regulate myeloid-biased hematopoietic stem cells via a histamine-dependent feedback loop. Cell stem cell, 21(6), 747-760.e7.
+ Open protocol
+ Expand
8

In Vitro T Cell Differentiation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Stocks of OP9 and OP9-DL1 stromal cells were a gift from J. C. Zúñiga-Pflücker. All experiments were performed in Opti-MEM with GlutaMAX (Gibco) containing 10% FCS (Gibco), 1% penicillin/streptomycin (Gibco), and 60mM 2-mercaptoethanol (Sigma-Aldrich) and maintained in a 37°C incubator (Thermo Scientific) with 5% CO2. Stromal cells were plated at 70% confluency. Before addition of lymphocytes, stromal cells were irradiated (1,500 rad) and culture media supplemented with murine IL-7 (25 ng ml−1; R&D Systems) and SCF (25 ng ml−1; BioLegend). Fetal liver lymphocytes were enriched for α4β7 by MACS and single-cell sorted onto 96 well plates containing stromal cells and cytokines as described above. Cultures were analyzed after 6 or 10 days of culture and only colonies with more than 10 CD45.2+ cells were considered.
Ishizuka I.E., Chea S., Gudjonson H., Constantinides M.G., Dinner A.R., Bendelac A, & Golub R. (2016). Single cell analysis defines the divergence between the innate lymphoid cell and lymphoid tissue inducer lineages. Nature immunology, 17(3), 269-276.
+ Open protocol
+ Expand
9

Culturing 293T, T-cells, and CD34+ Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
293T cells were maintained in Dulbecco’s Modification of Eagle’s Medium supplemented with glutamax, non-essential amino acids, penicillin/streptomycin and 10% fetal bovine serum ThermoFisher (Waltham, MA, USA). Cells were maintained at 37 °C and 5% CO2.
T-cells were grown in X-VIVO-20 (Lonza, Allendale, NJ, USA) with 10% AB serum (Valley Biomedical, Winchester, VA, USA), 300 IU of IL-2 and 5 ng/mL each of IL-7 and IL-15 (PeproTech, Rocky Hill, NJ, USA), N-Acetyl-l-Cysteine, penicillin/streptomycin, and Gluta-MAX-I each from ThermoFisher (Waltham, MA, USA).
CD34+ hematopoietic stem cells were cultured in StemSpan SFMII media containing 1 µM SR1 each from STEMCELL Technologies, Cambridge, MA, USA and human cytokines Flt-3 ligand (100 ng/mL), SCF (100 ng/mL), TPO (100 ng/mL), IL-6 (100 ng/mL) all from Biolegend, San Diego, CA, USA.
Osborn M.J., Lees C.J., McElroy A.N., Merkel S.C., Eide C.R., Mathews W., Feser C.J., Tschann M., McElmury R.T., Webber B.R., Kim C.J., Blazar B.R, & Tolar J. (2018). CRISPR/Cas9-Based Cellular Engineering for Targeted Gene Overexpression. International Journal of Molecular Sciences, 19(4), 946.
+ Open protocol
+ Expand
10

Hematopoietic Stem Cell Culture Conditions

Check if the same lab product or an alternative is used in the 5 most similar protocols
For growth assays, sorted CD34-CD150+LSK HSCs were cultured in S-Clone SF-O3 (Sanko Junyaku) supplemented with 0.1% BSA (093001; STEMCELL Technologies), 50 μM 2-ME (Sigma-Aldrich) and 1% penicillin/streptomycin/glutamine (Gibco). 20 ng/mL of recombinant mouse SCF (579706; BioLegend) and recombinant human TPO (763706; BioLegend) for HSC culture conditions and 10 ng/mL of SCF, TPO, recombinant mouse IL-3 (575506; BioLegend), and recombinant murine GM-CSF (315-03; PeproTech) for myeloid culture condition-1 were added to cultures. In the case of myeloid culture condition-2, sorted CD150+CD48-CD135-CD34-LSK HSCs, LSK cells, and GMPs were cultured in IMDM (Gibco) supplemented with 5% FBS, 50 μM 2-ME (Sigma-Aldrich), 1% penicillin/streptomycin/glutamine (Gibco), 1 mM sodium pyruvate (Gibco), and 0.1 mM MEM Non-Essential Amino Acids solution (Gibco). 25 ng/mL of SCF, TPO, recombinant human Flt3L (300-19; PeproTech) and recombinant murine IL-11 (220-11; PeproTech) and 10 ng/mL of IL-3 and GM-CSF were added to cultures.
For replating assays, LSK cells were plated in methylcellulose medium (Methocult M3234; STEMCELL Technologies) containing 20 ng/mL of SCF, TPO, IL-3, and GM-CSF.
Nakajima-Takagi Y., Oshima M., Takano J., Koide S., Itokawa N., Uemura S., Yamashita M., Andoh S., Aoyama K., Isshiki Y., Shinoda D., Saraya A., Arai F., Yamaguchi K., Furukawa Y., Koseki H., Ikawa T, & Iwama A. (2023). Polycomb repressive complex 1.1 coordinates homeostatic and emergency myelopoiesis. eLife, 12, e83004.
+ 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!