Anti c kit antibody

Manufactured by Santa Cruz Biotechnology

Sourced in United States

The Anti-c-kit antibody is a laboratory reagent used to detect and study the c-kit protein, which is a receptor tyrosine kinase involved in various cellular processes. This antibody can be used in techniques such as Western blotting, immunohistochemistry, and flow cytometry to identify and characterize the c-kit protein in biological samples.

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Lab products found in correlation

Facsaria, by BD (1 mentions) Anti cd45 antibody, by BD (1 mentions) Fortessa, by BD (1 mentions) Lymphocyte separation medium ficoll, by GE Healthcare (1 mentions) Annexin 5 fitc apoptosis detection kit, by Dojindo Laboratories (1 mentions) Anti p44 42 mapk erk1 2 antibody, by Cell Signaling Technology (1 mentions) Reverse transcription kit, by Promega (1 mentions) Anti histone h3 antibody, by Cell Signaling Technology (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Trizol, by Thermo Fisher Scientific (1 mentions) Cck 8, by Dojindo Laboratories (1 mentions) Enhanced chemiluminescence system, by Cytiva (1 mentions) Quality one image analysis software, by Bio-Rad (1 mentions) Anti β actin antibody, by Santa Cruz Biotechnology (1 mentions)

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Anti-c-kit (10 citations)

6 protocols using anti c kit antibody

Isolation and Culture of Murine Cardiac Progenitor Cells

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The isolation, characterization, and culture of
lin⁻/c-kit⁺ murine CPCs have been previously
described[11 (link)].
Lin⁻/c-kit⁺ CPCs (henceforth referred to simply
as CPCs) were isolated from GFP transgenic mice, RFP transgenic mice, and
wild-type (WT) mice (male, C57BL6, 8–10 weeks of age). Hearts were finely
minced and cultured to establish cell outgrowth cultures over ~7 days
using growth medium (F12 K medium supplemented with bFGF, LIF, and 10% FBS).
CPCs were isolated from the cell outgrowth of the explants by sequential sorting
procedures including depletion of mature hematopoietic lin⁺ cells and
sorting for c-kit with a specific anti-c-kit antibody (Santa Cruz) and magnetic
immunobeads (Miltenyi). To maximize the purity of the preparation, the c-kit
sorting procedure was repeated three consecutive times at 14-day intervals. The
lin⁻/c-kit⁺ cells were cultured, and the purity
of the sorted cells was confirmed quantitatively by flow cytometry and
immunofluorescent staining before use. In all studies, the CPCs used for cell
transplantation in vivo were passaged 4–6 times [11 (link)].

Guo Y., Nong Y., Li Q., Tomlin A., Kahlon A., Gumpert A., Slezak J., Zhu X, & Bolli R. (2020). Comparison of one and three intraventricular injections of cardiac progenitor cells in a murine model of chronic ischemic cardiomyopathy. Stem cell reviews and reports, 17(2), 604-615.

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Flow Cytometry Analysis of ckit+ CSCs

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For FACS analysis, ckit⁺CSCs were fixed in 4% paraformaldehyde for 15 minutes at room temperature and incubated with anti-CD45 antibody (BD Pharmingen) or anti-ckit antibody (Santa Cruz). Isotype control was performed (mouse polyclonal IgG; Santa Cruz Biotechnology). Specifically, cells were incubated with the primary antibody for 45 minutes at 37°C. Flow cytometry was performed with FACSAria (Becton Dickinson, San Jose, CA) instrument. Cellular debris and aggregates were gated out based on forward and side scatter. Gating on the signal of the nuclear stain DAPI was employed to exclude additional artifacts. Isotype-matched negative controls were utilized to define the threshold for each specific signal and establish the appropriate gate for positive cells. Data were analyzed with Kaluza Flow Cytometry Analysis software [76 (link)].

Puddighinu G., D’Amario D., Foglio E., Manchi M., Siracusano A., Pontemezzo E., Cordella M., Facchiano F., Pellegrini L., Mangoni A., Tafani M., Crea F., Germani A., Russo M.A, & Limana F. (2017). Molecular mechanisms of cardioprotective effects mediated by transplanted cardiac ckit+ cells through the activation of an inflammatory hypoxia-dependent reparative response. Oncotarget, 9(1), 937-957.

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Flow Cytometric Analysis of c-Kit+ Cells

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rCPCs were trypsinized and washed with PBS, incubated with 2 μg/10⁶ cells of anti‐c‐Kit antibody (Santa Cruz #sc‐5535) or IgG isotype (#sc‐3888) for 1 hour and washed with PBS, and then stained with Alexa Fluor 568 conjugated anti‐rabbit antibody (1:100, Thermo Fisher) for 1 hour. Dead cells were removed by 7‐Aminoactinomycin D staining. Flow cytometry analysis was carried out with Fortessa (BD Biosciences).

Mochizuki M., Lorenz V., Ivanek R., Della Verde G., Gaudiello E., Marsano A., Pfister O, & Kuster G.M. (2017). Polo‐Like Kinase 2 is Dynamically Regulated to Coordinate Proliferation and Early Lineage Specification Downstream of Yes‐Associated Protein 1 in Cardiac Progenitor Cells. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 6(10), e005920.

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Immunostaining of Pluripotent Stem Cells

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For in vitro staining, cells were plated on fibronectin-coated coverslips overnight, fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100 in PBS, and incubated with a rabbit polyclonal anti-c-kit antibody (cat #sc5535, Santa Cruz, USA, 1:100, 2h at 37°C), Oct3/4 (cat #ab18976, Abcam, USA, 1:100, 2h at 37°C), and Nanog (cat #sc33760, Santa Cruz, USA, 1:100, 2h at 37°C), followed by secondary anti-rabbit Alexa Fluor 488/594 antibodies (cat #A11001, A21207, all Invitrogen, USA, 1:800, 1h at 37°C); nuclei were counterstained by DAPI.

Dergilev K., Tsokolaeva Z., Makarevich P., Beloglazova I., Zubkova E., Boldyreva M., Ratner E., Dyikanov D., Menshikov M., Ovchinnikov A., Ageev F, & Parfyonova Y. (2018). C-Kit Cardiac Progenitor Cell Based Cell Sheet Improves Vascularization and Attenuates Cardiac Remodeling following Myocardial Infarction in Rats. BioMed Research International, 2018, 3536854.

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Chidamide-mediated Cell Apoptosis Analysis

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Chidamide was obtained from Shenzhen Chipscreen and prepared into 2 mM stock solution using Dimethyl sulfoxide (DMSO). The solution was preserved at −20 °C and thawed prior to use. RIPM1640 medium was purchased from Hyclone (USA). Fetal bovine serum (FBS) was purchased from Gibco (USA). CCK-8 was purchased from Dojindo (Japan). Annexin V-FITC apoptosis detection kit was manufactured by Dojindo. Anti-histone H3 antibody, anti-acetyl-histone H3 antibody, anti-p44/42MAPK (ERK1/2) antibody, and anti-Pp44/42MAPK (ERK1/2) antibody were purchased from Cell Signaling (USA). Anti-AML1/RHD antibody was purchased from Calbiochem (USA). Anti-C-KIT antibody, β-acting, and HRP-labeled secondary antibodies were purchased from Santa Cruz (USA). Lymphocyte separation medium (Ficoll) was purchased from GE Healthcare (USA). Trizol was purchased from Ambion (USA). Reverse transcription kit was purchased from Promega (USA). KAPA SYBR FAST q-PCR Master Kit was purchased from KAPA (USA).

Liu J., Lv N., Zhou L., Li Y, & Yu L. (2020). Chidamide inhibits t(8;21) AML cell proliferation and AMK1/ETO and C-KIT expression by inhibiting ERK1/2 signaling pathway. Translational Cancer Research, 9(2), 827-839.

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Western Blot Analysis of c-kit, ABCG2, and β-Actin

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Cells were collected in standard sodium dodecyl sulfate (SDS) sample buffer. Protein concentrations were determined using the Bradford method, and protein extracts (20 mg/lane) were separated on a 10% SDS-polyacrylamide gel, transferred to a nitrocellulose membrane, and probed with 1:200 anti-c-kit antibody (Santa Cruz Biotechnology, Inc., CA, USA), 1:1000 anti-ABCG2 antibody (Abcam Inc., Cambridge, MA, USA), or 1:1000 anti-b-actin antibody (Santa Cruz). Proteins were visualized using an enhanced chemiluminescence system (Amersham, UK) and were analyzed using the QualityOne Image Analysis software (Bio-Rad, Hercules, CA, USA). The experiments were repeated three times.

Zheng J.C., Qi S.Q., Yang L., Ma Y.Y., Dong K.R., Zhu H.T., Yang S.B., Xu T., Zheng S, & Xiao X.M. (2015). Effects of bisphenol A on decreasing the percentage and promoting the growth of stem cell-like cells from SK-N-SH human neuroblastoma cells. Genetics and molecular research : GMR, 14(2).

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