Anti mouse cd45 antibody

Manufactured by BioLegend

The Anti-mouse CD45 antibody is a laboratory reagent used for the identification and characterization of mouse immune cells. CD45 is a protein tyrosine phosphatase expressed on the surface of all hematopoietic cells. This antibody can be used in flow cytometry and other immunoassays to detect and analyze CD45-positive cells.

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Lab products found in correlation

Quadromacs column separation kit, by Miltenyi Biotec (1 mentions) Anti mouse ep cam antibody, by BioLegend (1 mentions) Dead cell removal kit, by Miltenyi Biotec (1 mentions) Trichlorosilane, by Merck Group (1 mentions) Rat igg2b, by BioLegend (1 mentions) 1 step ultra tmb elisa, by Thermo Fisher Scientific (1 mentions) Biotin 5 fluorescein conjugate, by Merck Group (1 mentions) Hrp anti mouse igg, by BioLegend (1 mentions) Penicillin streptomycin, by Corning (1 mentions) Alexa fluor 488 streptavidin, by BioLegend (1 mentions) Biotinylated anti mouse igg antibody, by Jackson ImmunoResearch (1 mentions) Carboxylated polybeads, by Polysciences (1 mentions) Superblock t20 blocking buffer, by Thermo Fisher Scientific (1 mentions) Polydimethylsiloxane pdms, by Dow (1 mentions) Fetal bovine serum (fbs), by Atlanta Biologicals (1 mentions) Macs column, by Miltenyi Biotec (1 mentions) Amicon ultra centrifugal filter, by Merck Group (1 mentions) 7 aad, by BioLegend (1 mentions) Anti mouse cd3 antibody, by BioLegend (1 mentions) Facsaria 2, by BD (1 mentions)

4 protocols using anti mouse cd45 antibody

Isolation and Purification of Liver Cell Populations

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Parenchymal and non-parenchymal cells from mouse liver were isolated by a 2-step collagenase perfusion (16 (link)). The parenchymal cell fraction, which consists mostly of hepatocytes, also contains hepatic stellate cells due to their close physical association with hepatocytes in vivo. Positive selection of CD45+ cells and EpCAM+ cells from non-parenchymal cells was performed using the QuadroMACS column separation Kit (Miltenyi Biotech, Cambridge, MA) (10 (link)). Cells were then purified using the Dead Cell Removal Kit (Miltenyi Biotech). Anti-mouse CD45 antibody, anti-mouse EpCAM antibody (Biolegend, San Diego, CA) and microbeads were used according to the manufacturer’s instructions. To assess the purity of EpCAM+ cells, low expression of CD68 (Kupffer cells) and Tie2 (endothelial cells) was confirmed by qRT-PCR.
Additional methods are available in the online Supplementary Methods.

Okabe H., Yang J., Sylakowski K., Yovchev M., Miyagawa Y., Nagarajan S., Chikina M., Thompson M., Oertel M., Baba H., Monga S.P, & Nejak-Bowen K.N. (2016). Wnt signaling regulates hepatobiliary repair following cholestatic liver injury in mice. Hepatology (Baltimore, Md.), 64(5), 1652-1666.

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Histological Analysis of DRG Tissues

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DRG was fixed in formalin and embedded in paraffin. Tissue sections of 5μm thickness were stained with hematoxylin and eosin (H&E). For immunohistochemistry staining, tissue sections were stained with anti- mouse CD45 antibody (Biolegend). The slides were captured at 20 x magnification using Olympus camera.

Liang Y., Yang K., Guo J., Wroblewska J., Fu Y.X, & Peng H. (2015). Innate lymphotoxin receptor mediated signaling promotes HSV-1 associated neuroinflammation and viral replication. Scientific Reports, 5, 10406.

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Fabrication and Characterization of Biotinylated Surfaces

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Tetramethylammonium hydroxide (TMAOH), streptavidin, poly(ethylene glycol)bis(amine) (PEG-diamine, average M_n 3,400), biotin (5-fluorescein) conjugate, and trichlorosilane (from Sigma-Aldrich); Dynabead™ M-280 streptavidin, ethyl-3-(3-(dimethylamino)propyl)carbodiimide (EDC), hydroxysulfosuccinimide (sulfo-NHS), bis(sulfosuccinimidyl) suberate (BS3), SuperBlock T20 blocking buffer, Glutamax, and 1-Step Ultra TMB-ELISA (from Thermo Fisher Scientific); fetal bovine serum (from Atlanta Biologicals); penicillin-streptomycin (from Cellgro); DMEM media (from Mediatech); polydimethylsiloxane (PDMS) (from Dow Corning); 10 μm fluorescent microbeads (from Bangs Laboratory); carboxymethyl (CM) dextran (from PK chemicals A/S); 3 μm carboxylated polybeads (from Polyscience); Biotinylated anti-mouse IgG antibody (from Jackson ImmunoResearch); HRP anti-mouse-IgG, Alexa Fluor 488 streptavidin, anti-mouse CD45 antibody, Rat IgG2b (from BioLegend); MACS column (from Miltenyi Biotec); Amicon Ultra centrifugal filters (from Merck Millipore) were used as received. All chemicals were of analytical grade and used without purification.

Park K.S., Kim H., Kim S., Lee K., Park S., Song J., Min C., Khanam F., Rashu R., Bhuiyan T.R., Ryan E.T., Qadri F., Weissleder R., Cheon J., Charles R.C, & Lee H. (2017). Nano-Magnetic System For Rapid Diagnosis Of Acute Infection. ACS nano, 11(11), 11425-11432.

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Dissociation and FACS of Appendage Tissue

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Freshly dissected tissue from the rim of healing appendages were manually dissociated with scissors and then incubated with liberase (Roche) for 60 min at 37°C. After dissociation, cells were washed in PBS and resuspended in FACS buffer (HBSS, 0.2% BSA, 1% HEPES). 7-AAD (BioLegend) was added to exclude dead cells. FACS was performed on a FACS Aria II (BD Biosciences). Standard FACS procedures were performed; all positive sorting gates were based on negative gates, and compensation was carried out to remove spectral overlap. Sorted cells were immediately harvested for RNA purification and real-time RT–PCR analysis. The following primary antibodies were used: anti-mouse CD3 antibody (BioLegend) and anti-mouse CD45 antibody (BioLegend).

Leung T.H., Snyder E.R., Liu Y., Wang J, & Kim S.K. (2015). A cellular, molecular, and pharmacological basis for appendage regeneration in mice. Genes & Development, 29(20), 2097-2107.

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