Whole bone marrow (WBM) cells were isolated from femurs and tibias by grinding the bones in HSC buffer. RBCs were lysed using ACK lysing buffer (Lonza). Total number of WBM cells was counted with a Coulter counter (Beckman Coulter). Three million WBM cells were blocked with a rat anti-mouse CD16/32 antibody (BD Pharmingen) and stained with PE-Cy5 conjugated anti-mouse CD3e (clone: 145-2C11), PE-Cy5 conjugated anti-mouse CD4 (clone: GK1.5), PE-Cy5 conjugated anti-mouse CD8 (clone: 53-6.7) PE-Cy5 conjugated anti-mouse B220 (clone: RA3-6B2), PE-Cy5 conjugated PE-Cy5 conjugated anti-mouse CD11b (clone: M1/70), PE-Cy5 conjugated anti-mouse Gr-1 (clone: RB6-8C5), PE-Cy5 conjugated anti-mouse Ter119 (Ly-76), PE conjugated anti-mouse Sca1 (clone:D7), APC conjugated anti-mouse cKit (clone: 2B8), FITC conjugated anti-mouse CD48 (clone: HM48-1) (eBioscience) and Brilliant Violet 421 conjugated anti-mouse CD150 (clone: TC15-12F12.2) antibodies (BioLegend). All antibodies were diluted 1:400. Dead cells were excluded by staining with 7-AAD (BD Pharmingen). Data were collected from 1 million single cells by FACSCanto (BD Pharmingen) and analyzed by FlowJo (Tree Star, Inc) without knowledge of the genotype or treatment by a single observer (CLL).
Pe conjugated anti mouse sca1
Manufactured by Thermo Fisher Scientific
Sourced in United States
The PE conjugated anti-mouse Sca1 is a laboratory reagent used for the detection and analysis of the Sca1 marker on mouse cells. It is a monoclonal antibody that is conjugated with the fluorescent dye phycoerythrin (PE), which allows for the visualization and quantification of Sca1-positive cells using flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Facscanto, by BD (3 mentions)
Rat anti mouse cd16 32 antibody, by BD (3 mentions)
Pe cy5 conjugated anti mouse cd4, by Thermo Fisher Scientific (3 mentions)
Pe cy5 conjugated anti mouse cd3e, by BD (3 mentions)
Pe cy5 conjugated anti mouse cd8, by Thermo Fisher Scientific (3 mentions)
Pe cy5 conjugated anti mouse b220, by Thermo Fisher Scientific (3 mentions)
Pe cy5 conjugated anti mouse gr 1, by Thermo Fisher Scientific (3 mentions)
Apc conjugated anti mouse ckit, by Thermo Fisher Scientific (3 mentions)
7 aad, by BD (2 mentions)
Coulter counter, by Beckman Coulter (2 mentions)
Ack lysing buffer, by Lonza (2 mentions)
Fitc conjugated anti mouse cd48, by Thermo Fisher Scientific (2 mentions)
Brilliant violet 421 conjugated anti mouse cd150, by BioLegend (2 mentions)
Pe cy5 conjugated anti mouse cd11b, by Thermo Fisher Scientific (1 mentions)
Lsm 710 confocal microscope, by Zeiss (1 mentions)
Cm3050s cryostat, by Leica (1 mentions)
Fitc conjugated anti mouse cd45, by Thermo Fisher Scientific (1 mentions)
4 protocols using pe conjugated anti mouse sca1
1
Multiparametric Flow Cytometry Analysis of Murine Hematopoietic Cells
2
Murine Hematopoietic Stem Cell Immunophenotyping
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
WBM cells were isolated from femurs and tibias by grinding the bones in HSC buffer. RBCs were lysed using ACK lysing buffer (Lonza). Total number of WBM cells was counted with a Coulter counter (Beckman Coulter). Three million WBM cells were blocked with a rat anti-mouse CD16/32 antibody (BD Pharmingen) and stained with PE-Cy5 conjugated anti-mouse CD3e (clone: 145-2C11), PE-Cy5 conjugated anti-mouse CD4 (clone: GK1.5), PE-Cy5 conjugated anti-mouse CD8 (clone: 53-6.7) PE-Cy5 conjugated anti-mouse B220 (clone: RA3-6B2), PE-Cy5 conjugated anti-mouse CD11b (clone: M1/70), PE-Cy5 conjugated anti-mouse Gr-1 (clone: RB6-8C5), PE-Cy5 conjugated anti-mouse Ter119 (Ly-76), PE-conjugated anti-mouse Sca1 (clone:D7), APC conjugated anti-mouse cKit (clone: 2B8), FITC conjugated anti-mouse CD48 (clone: HM48-1; eBioscience) and Brilliant Violet 421 conjugated anti-mouse CD150 (clone: TC15-12F12.2; BioLegend) antibodies. All antibodies were diluted 1:400. Dead cells were excluded by staining with 7AAD (BD Pharmingen). Data were collected from 1 million single cells by FACSCanto (BD Pharmingen) and analysed by FlowJo (Tree Star, Inc.) without knowledge of the genotype or treatment by a single observer (C.L.L.).
3
Multicolor Immunofluorescence Staining of Bone Marrow
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Frozen BM sections were prepared and immunostained according to the Kawamoto method, as previously reported [28 (link)]. Dissected femurs were embedded in Super Cryoembedding Medium (SCEM) and fixed using dry ice and hexane. Bones were cryosectioned (7 μm sections) using cryofilms and a CM3050s cryostat (Leica Biosystems Nussloch GmbH, Biberach, Germany). Immunofluorescence data were obtained and analyzed with a LSM710 confocal microscope (Carl Zeiss Japan, Tokyo, Japan). The markers and antibodies were as follows: Hoechst (a DNA marker used to detect cell nuclei), FITC-conjugated anti-mouse stem cell antigen 1 (Sca-1), phycoerythrin (PE)-conjugated anti-mouse cluster of differentiation (CD) 45, PE-conjugated anti-mouse TER-119, APC-conjugated anti-mouse platelet-derived growth factor receptor α (PDGFRα/CD140a) (eBioscience, San Diego, CA, USA), PE-conjugated anti-mouse Sca-1, and FITC-conjugated anti-pimonidazole hydrochloride.
4
Comprehensive BM Cell Immunophenotyping
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
Revolutionizing how scientists
search and build protocols!