For autophagic flux analysis, retroviruses expressing pBabe-GFP-cherry-LC3 (construct was a gift from Dr. Debnath’s lab at UCSF) were made using GP2-293 cells transfected with pBabe GFP-cherry-LC3 and pVSV-G plasmids using TransIT-LT1 (Mirus) transfection reagent. After puromycin selection, cells were sorted in a Moflo XDP 100 cell sorter (Beckman Coulter, Inc.) for the high (top 15%) fluorescent population. Normal or GFP-cherry-LC3 expressing cells were stained with V450 mouse anti-human CD44 (BD Bioscience, 561292) or Alexa Fluor 647 mouse anti-human CD24 (BD Bioscience, 561644) or their corresponding isotypes (V450 mouse IgG2b, k Isotype control and Alexa Fuor 647 mouse IgG2a, k Isotype control (BD Biosciences, catalog numbers 560374 and 557715), washed, resuspended in 0.5% BSA/ PBS and analyzed in a Gallios 561 (Beckman Coulter) flow cytometer.
V450 mouse igg2b
Manufactured by BD
Sourced in United States
The V450 mouse IgG2b is a laboratory reagent used for flow cytometry applications. It is a fluorescent-conjugated antibody that specifically binds to the IgG2b isotype of mouse immunoglobulins.
Automatically generated - may contain errors
Lab products found in correlation
Gallios 561, by Beckman Coulter (1 mentions)
V450 mouse anti human cd44, by BD (1 mentions)
Alexa fluor 647 mouse anti human cd24, by BD (1 mentions)
Apc mouse igg1, by BD (1 mentions)
V450 mouse anti human cd14, by BD (1 mentions)
7 aminoactinomycin d 7 aad, by Thermo Fisher Scientific (1 mentions)
Facsmelody, by BD (1 mentions)
Facsaria fusion, by BD (1 mentions)
2 protocols using v450 mouse igg2b
1
Autophagic Flux Analysis with GFP-Cherry-LC3
2
Flow Cytometric Analysis of Neutrophil and Monocyte Differentiation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After collection, cells were washed with PBS twice at room temperature. Cells under neutrophil differentiation were then incubated with 7-aminoactinomycin D (7-AAD) (ThermoFisher Scientific, Waltham, MA, USA, cat. No. A1310) and PE-conjugated mouse anti-human CD11b (clone ICRF44) (BD Biosciences, San Jose, CA, USA, cat. No. 557321) or PE-conjugated isotype control mouse mAb (clone: MOPC-21) (Biolegend, San Diego, CA, USA, cat. No. 400112) for 40 min at 4 °C in the dark. Samples were analyzed by BD FacsAria Fusion with a 100-micron nozzle at 4 °C. Cells under monocyte differentiation were incubated with V450 mouse anti-human CD14 (BD Biosciences, San Jose, CA, USA, cat. No. 560349) and adenomatous polyposis coli (APC) mouse anti-human CD71 (BD Biosciences, San Jose, CA, USA, cat. No. 551374) or V450 mouse IgG2b (BD Biosciences, San Jose, CA, USA, cat. No. 560374) and APC mouse IgG1 (BD Biosciences, San Jose, CA, USA, cat. No. 555751) for isotype control. Samples were analyzed by BD FACSMelody at 4 °C. All data were analyzed with FlowJo software v10.6.1 downloaded from https://www.flowjo.com/ .
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!