CTC isolation was conducted in the mouse model of orthotopic metastatic HCC. Whole blood was withdrawn by heart puncture at euthanasia and immediately transferred into a 1.5‐mL EDTA disposable tube. The blood was immediately processed for CTC enrichment by Ficoll gradient, adapting the manufacturer’s protocol for the amount of blood collected. The blood was diluted in 2 mL of PBS and carefully layered on 3 mL of Ficoll Paque reagent (GE Healthcare). After centrifugation (400g at room temperature according to the manufacturer’s instructions), the PBMC layer was collected. If evident red cell contamination was detected at macroscopic examination, a red cell lysis step was added by using a commercial red cell lysis buffer (e‐Bioscience). After two washing steps with 2.5% fetal bovine serum–PBS, the cells were immediately stained with a fluorescent anti‐mouse monoclonal cluster of differentiation (CD)45 antibody (AF700CD45; BioLegend), according to the manufacturer’s instructions, and then analyzed by flow cytometry using DIVA software. We identified miRNA‐517 cells by flow cytometry as GFP‐positive cells.
Af700cd45
Manufactured by BioLegend
AF700CD45 is a fluorescent-conjugated monoclonal antibody that specifically binds to the CD45 protein, a common leukocyte antigen expressed on the surface of various immune cells.
Automatically generated - may contain errors
Lab products found in correlation
Red cell lysis buffer, by Thermo Fisher Scientific (1 mentions)
Ficoll paque reagent, by GE Healthcare (1 mentions)
Pacific blue ifnγ, by BioLegend (1 mentions)
Anti mouse pd 1, by BioXCell (1 mentions)
Cd62l pe cy7, by BioLegend (1 mentions)
Percp cy5.5 pd 1, by BioLegend (1 mentions)
Fixation permeabilization kit, by BD (1 mentions)
Cd44 apc cy7, by BioLegend (1 mentions)
Cd8a apc, by BioLegend (1 mentions)
Live dead dye, by Thermo Fisher Scientific (1 mentions)
Cd4 fitc, by BioLegend (1 mentions)
Brefeldin a golgiplug, by BD (1 mentions)
Bv605 tnfα, by BioLegend (1 mentions)
Anti mouse ctla 4, by BioXCell (1 mentions)
Collagenase type 1, by Thermo Fisher Scientific (1 mentions)
Dnase 1, by Roche (1 mentions)
Fc block, by BD (1 mentions)
Easysep dead cell removal annexin 5 kit, by STEMCELL (1 mentions)
Facsaria 2, by BD (1 mentions)
3 protocols using af700cd45
1
Isolation and Identification of Circulating Tumor Cells in Metastatic HCC Mouse Model
2
Peptide Library Synthesis and Liposome Formulation
3
CD45+ Cell Isolation from Paw Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Following paw tissue digestion, as described above, live cells were obtained using EasySepTM Dead Cell Removal (Annexin V) kit (STEM CELL) according to manufacturer's instruction. Cells were incubated with AF700 CD45 (Biolegend) at a dilution of 1:100 in DPBS−/− with 0.02 % BSA for 30 minutes at 4°C. Non-specific staining was blocked with TruStain X. Cells were suspended in DPBS−/− with 0.02 % BSA and the BD FACS Aria II was used sort for CD45 positive cells, which were collected in DPBS containing 0.1 % BSA for single cell sequencing.
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!