The fresh in-situ HCC xenograft tissues were perfused at a flow rate of 10 mL/min with Gey's balanced salt solution (GBSS) for 10 min, followed by 100 mL of 0.12% pronase E (Roche) dissolved in GBSS for another 10 min [19 (link), 30 (link)]. The HCC tissues were then excised, dissected and incubated for 30 min with continuous shaking, with 0.04% pronase E, 0.05% collagenase and 0.002% DNase I (Sigma) in 100 mL GBSS. After digestion, the cell suspension was passed through a 0.22-μm mesh and centrifuged at 500 × g for 10 min. Subsequently, cells were purified with 8% Nycodenz (Sigma) gradient centrifugation. The resulting monocytes were then sorted by flow cytometry with anti-CD11c antibody (Abcam, Suzhou, China). Purified DCs were then generated via culturing in 6-well tissue-culture plates (Costar) plus 50 ng/mL GM-CSF and 10 ng/mL IL-4 (1,000 U/mL, R&D systems) for 5 days in RPMI 1640 FCS medium (no antibiotic). On day-5, DCs were determined by flow cytometry with anti-CD11c antibody (over 90% positive rate).
Anti cd11c antibody
Manufactured by Abcam
Sourced in United Kingdom, United States
Anti-CD11c antibody is a laboratory reagent used in research applications. It is designed to specifically detect the CD11c protein, which is expressed on the surface of certain immune cells such as dendritic cells and macrophages. This antibody can be used in techniques like flow cytometry, immunohistochemistry, and Western blotting to identify and study these cell types.
Automatically generated - may contain errors
Lab products found in correlation
Collagenase, by Merck Group (1 mentions)
Nycodenz, by Merck Group (1 mentions)
6 well tissue culture plate, by Corning (1 mentions)
Pronase e, by Roche (1 mentions)
Pronase e, by Merck Group (1 mentions)
Dnase 1, by Merck Group (1 mentions)
Gm csf, by R&D Systems (1 mentions)
Interleukin 4 (il 4), by R&D Systems (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Cd80 fitc, by Thermo Fisher Scientific (1 mentions)
Cd86 pe, by Thermo Fisher Scientific (1 mentions)
Interleukin 4 (il 4), by Thermo Fisher Scientific (1 mentions)
Recombinant gm csf, by Thermo Fisher Scientific (1 mentions)
Lps from escherichia coli 026 b6, by Merck Group (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Anti f4 80, by Abcam (1 mentions)
Anti cd206 antibody, by Abcam (1 mentions)
Anti f4 80 antibody, by Abcam (1 mentions)
Flow cytometry, by Beckman Coulter (1 mentions)
6 protocols using anti cd11c antibody
1
Isolation of Dendritic Cells from HCC Xenograft Tissues
2
Apoptosis Signaling Pathway Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Rabbit polyclonal anti-Bax (Cat. No.A19684), anti-Caspase 3(Cat. No. A11319) and anti-Actin (Cat. No. AC026) antibodies were purchased from Abclonal (Wuhan, China), and rabbit polyclonal anti-Bcl2 antibody (Cat. No. 12789-1-AP) was obtained from Proteintech (Wuhan, China). Anti-CD11c antibody (Cat.No.ab52632) was obtained from Abcam (Cambridge, United Kingdom). Arginase inhibitor 1 (Cat.No.1345808-25-4) was obtained from MedChemExpress (New Jersey, United States).
3
Dendritic Cell Differentiation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RPMI 1640 medium, streptomycin, and penicillin were purchased from Invitrogen (Grand Island, NY, USA). Fetal bovine serum (FBS) was purchased from Hyclone (Thermo, Melbourne, Australia). Recombinant GM-CSF and IL-4 were purchased from Peprotech (Rocky Hill, NJ, USA). LPS (from Escherichia coli 026:B6) was obtained from Sigma-Aldrich (St Louis, MO, USA). The fluorescent-labeled anti-mouse CD40-PE, CD80-FITC, and CD86-PE mAbs were purchased from eBioscience (San Diego, USA). The anti-CD11c antibody and anti-influenza virus nucleoprotein antibody (FITC-conjugated) were purchased from Abcam (MA, USA).
4
Isolation of Primary Alveolar Macrophages
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary alveolar macrophages were isolated from rodents following established protocols with minor modifications [58 (link)]. Briefly, mice and rats were killed and the lungs were carefully removed. The lung was cannulated via the trachea and BAL was performed using 10×5 ml ice-cold PBS. The lavage fluid was centrifuged at 400 g for 5 min and the pelleted cells were resuspended in PBS. Cell viability was determined by trypan blue exclusion. The cells were cultured in Ham’s F-12K medium supplemented with 15% FBS for 45 min at 37 °C and the non-adherent cells was washed away with PBS. The adherent cells were recovered and diluted as needed. Alveolar macrophages were identified as F4/80+ and CD11c+ cells by flow cytometry using anti-F4/80 (Abcam) and anti-CD11c antibody (Abcam) [59 (link)].
5
Evaluating M1 and M2 Macrophage Polarization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To assess the M1 or M2 polarization of macrophages, we carried out flow cytometric detection of M1 or M2 macrophages from ischemic lower limb tissue of mice. Cultured macrophages were isolated and harvested at a single-cell level, and surface markers were analyzed. The isolated cells were subsequently incubated with fluorescein isothiocyanate (FITC)-conjugated or phycoerythrin (PE)-conjugated monoclonal antibodies. The anti-F4/80 antibody (Abcam, Cambridge, MA, USA), anti-CD11c antibody (Abcam), and anti-CD206 antibody (Abcam) were conjugated monoclonal antibodies. M1 macrophages were identified on the basis of being F4/80 and CD11c positive, while M2 macrophages were CD206 positive. F4/80 and CD11c double-positive M1 macrophages as well as F4/80 and CD206 double-positive M2 macrophages from ischemic lower limb tissues of mice were detected by flow cytometry (Beckman Coulter, Fullerton, CA, USA) at day 1, 3, 7, and 14 after the surgical procedure, respectively. The data were analyzed using FlowJo software (Tree Star, Inc., Ashland, OR, USA). The ratio of M1 to M2 macrophages was observed.
6
Visualizing SARS-CoV-2 Spike Protein and Dendritic Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Local skin tissues from immunized mice were immediately frozen in liquid nitrogen. The tissue sections were embedded, sliced, fixed, and blocked using 5% bovine serum albumin (BSA). For detection of the viral antigen, the sections were sequentially incubated with a primary mouse anti-SARS-CoV-2 spike antibody (SinoBiological Co., Ltd) and an AlexaFluor 647-conjugated goat anti-mouse IgG secondary antibody (Invitrogen, Carlsbad, CA, USA). Dendritic cells were detected using anti-CD11c antibody (Abcam, Cambridge, UK) and Alexa Fluor® 488 goat anti-rabbit IgG secondary antibody (Invitrogen). The cell nuclei were detected using DAPI. Fluorescence was visualized and analyzed using a confocal microscope (TCS SP2, Leica).
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!