Expression of CTSB on the surface of YPK2 and YPK2-Lm cells was analyzed using flow cytometry. Dissociated cells were suspended in PBS with 2% FBS (106 cells/100 µl) and then incubated with each antibody at 4°C for 30 min. The antibodies for this assay were rat anti-human CD44v9 (1:50; clone RV3; cat. no. LKG-M003; Cosmo Bio Co., Ltd.), anti-calreticulin (CALR; 1:50; cat. no. ab196159; Abcam) and anti-CTSB (1:20; cat. no. ab58802; Abcam). The mouse FITC-conjugated anti-rat IgG2a secondary antibody (1:10; cat. no. 11-4811-85; eBioscience; Thermo Fisher Scientific, Inc.) was used for the anti CD44v9 primary antibody. The anti-CTSB and cognate isotype control antibodies were conjugated with APC using an APC conjugation kit (cat. no. ab201807; Abcam) according to the manufacturer's instructions. Rat IgG2a κ Isotype Control (cat. no. 14-4321-82; eBioscience; Thermo Fisher Scientific, Inc.), rabbit IgG Isotype Control Alexa Fluor 647 (cat. no. ab199093; Abcam) and mouse IgG2a κ monoclonal Isotype Control (cat. no. ab18415; Abcam) were used as negative controls for anti-CD44v9, anti-CRT Alexa Fluor 647 and anti-CTSB with corresponding dilution factors. Flow cytometry analysis was performed using a FACS ARIA-III (BD Biosciences) and a MACSQuant analyzer version 2.4 (Miltenyi Biotec GmbH).
Anti crt alexa fluor 647
Manufactured by Abcam
Sourced in United States
Anti-CRT Alexa Fluor 647 is a fluorescently labeled antibody targeting the CRT (calreticulin) protein. Alexa Fluor 647 is the fluorescent dye conjugated to the antibody. This product can be used for various applications that require detection of the CRT protein, such as immunofluorescence and western blotting.
Automatically generated - may contain errors
Lab products found in correlation
Anti ctsb, by Abcam (1 mentions)
Facsaria 3, by BD (1 mentions)
Anti cd31 apc, by Thermo Fisher Scientific (1 mentions)
Anti mhcii efluor450, by Thermo Fisher Scientific (1 mentions)
Anti cd11b pe, by Thermo Fisher Scientific (1 mentions)
Anti cd45 pacific blue, by Thermo Fisher Scientific (1 mentions)
Ii type collagenase, by Merck Group (1 mentions)
Anti f4 80 pe, by Thermo Fisher Scientific (1 mentions)
Anti ly6c fitc, by Thermo Fisher Scientific (1 mentions)
Trypsin, by Merck Group (1 mentions)
2 protocols using anti crt alexa fluor 647
1
Flow Cytometric Analysis of CTSB Expression
2
Muscle-Derived Immune Cell Profiling
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Using 0.2% II type collagenase (Sigma, USA), inflamed TA muscles were collected and digested for 40 min at the condition of 37 °C. In vivo, the single cell suspension obtained from muscle homogenate was blocked. In vitro, cultured cells were digested with Trypsin (Sigma, USA), resuspended in ice cold PBS to obtain the single cell suspension. The following fluorescent antibodies were used: anti-CD45-Pacific Blue, anti-F4/80-PE, anti-CD11b-PE, anti-MHC-II-eFluor 450, anti-Ly6C-FITC, anti-CX3CR1-APC, anti-CD206-eFluor 700, anti-Bcl3-FITC, anti-CD31-APC, anti-IL-10-FITC, rabbit anti-p-STAT3-FITC, the antibodies above were purchased from ThermoFisher and their dilution ratios were 1:100; Other antibodies involved anti-Vav1-FITC (1:100, Biorbyt, USA), anti-Rac1-GTP-FITC (1:100, Proteintech, USA), anti-Tunel-FITC (5:50, Yeason, China), anti-Annexin-V-APC (5:100, Sigma, USA), anti-CRT-Alexa Fluor 647 (1:100, Abcam, UK), anti-PKH67-Alexa Fluor 647 (1:250, Zeye, China), anti-CD36-Alexa Fluor 700 (1:100, eBioscience, USA), and anti-PPARγ-FITC (1:100, Abcam, UK). To analyze the labeled cells, FACSAria II cell sorter with FlowJo software (BD Biosciences, USA) were used.
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