KTC1 and U87 cells were treated as indicated at 37℃. The following fluorophore-conjugated antibodies were used: CD31 (BD; WM59), PD-L1 (BD; MI5), PD-L2 (BD; MI18), and VISTA (R&D; 730804). Anti-mouse CD16/CD32 (BD; mouse Fc blocker, clone 2.4G2) were used as the blocking reagent to reduce the non-specific binding of the antibodies. For flow cytometry analysis of in vivo tumor experiments, samples were stained with Fixable Viability Stain 510 or 780 (BD Horizon) and fluorescent dye-conjugated antibodies anti-mouse CD45 (HI30), CD3 (BD; SK7), CD4 (Biolegend; RPA-T4), CD8 (Biolegend; 53–6.7), CD11B (BD; M1/70), CD11C (BD; B-Iy6), CD19 (BD; 1D3), CD68 (BD; FA/11), Gr1 (BD; RB6-8C5), IFNγ (Biolegend; XMG1.2), and PD-L1 (BD; MI5). For FoxP3 detection, cells were stained using Transcription Factor Staining Set (BD) and anti-human FoxP3 antibody (236A/E7). For intracellular staining of interferon (IFN)γ, Cytofix/Cytoperm solution (BD) was added before fixation and permeabilization. We performed the acquisition with FACS LSRII (BD Biosciences) and data analysis in FlowJo software (FlowJo LLC, Ashland, Oregon, USA). Flow cytometry graphs shown in the results section were representative data from at least three independent experiments.
Pd l1
Manufactured by BD
Sourced in United States, Germany
The PD-L1 is a laboratory equipment product designed for use in research and development settings. It serves as a tool for the detection and analysis of the programmed death-ligand 1 (PD-L1) protein, which plays a role in immune system regulation. The product's core function is to facilitate the measurement and quantification of PD-L1 expression levels in various biological samples, enabling researchers to study its role in cellular processes and potential therapeutic applications.
Automatically generated - may contain errors
Lab products found in correlation
Pd l2, by BD (6 mentions)
Ctla 4, by BD (6 mentions)
F4 80, by BD (5 mentions)
Cd11b, by BD (4 mentions)
Ifn γ, by BD (4 mentions)
Facscalibur, by BD (4 mentions)
Hla dr, by BD (4 mentions)
Facscanto 2, by BD (4 mentions)
Granzyme b, by BD (3 mentions)
T bet, by BD (3 mentions)
Tnf α, by R&D Systems (3 mentions)
Fluorochrome conjugated anti human eomes mab, by Thermo Fisher Scientific (3 mentions)
Ifn α, by R&D Systems (3 mentions)
Cytoflex, by Beckman Coulter (3 mentions)
Pd l1, by Thermo Fisher Scientific (3 mentions)
Tim 3, by BD (3 mentions)
Recombinant human il 2, by R&D Systems (2 mentions)
Cd40l, by BD (2 mentions)
Tcrαβ, by BD (2 mentions)
Interleukin 4 (il 4), by R&D Systems (2 mentions)
44 protocols using pd l1
1
Flow Cytometry Analysis of Tumor Immune Cells
2
Multiparametric Phenotyping of T Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fluorochrome conjugated anti-human mAb specific to HLA-A2, CD3, CD8, CD28, CD38, CD40L, CD69, ICOS, TCRαβ, CCR7, CD45RO, CD80, CD86, ICOS ligand, IFN-γ, CTLA-4, PD-1, PD-L1, PD-L2, T-bet, granzyme B or Akt were purchased from Becton Dickinson (San Diego, CA). Fluorochrome conjugated anti-human Eomes mAb was purchased from eBioscience (San Diego, CA). Recombinant human IL-2, IL-4, IFN-α and TNF-α were purchased from R&D Systems, and human GM-CSF was obtained from Immunex (Seattle, WA). Lenalidomide (Celgene, Summit, NJ) was reconstituted in 1% DMSO and was used at a final concentration of 5 μM to treat either XBP1-CTL or various tumor cell lines.
3
Multiparametric Phenotyping of T Cells
4
Multiparametric Flow Cytometry Panel
5
Multiparametric Flow Cytometry Analysis of Immune Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Rat spleens and HCC tumors were harvested after euthanization. Spleens and HCC cells were homogenized separately and stained with the following antibodies: CD3-PE, CD4-PEcy7, CD8-FITC, CD-25-APCcy7, FoxP3-PECF594, PD-1, and PD-L1 (Becton Dickinson, Franklin Lakes, NJ, USA). BD LSRFortessa 6-Laser flow cytometer (Becton Dickinson, Franklin Lakes, NJ, USA). Data analysis was conducted with Flowjo software (Flowjo LLC, Becton Dickinson, Ashland, OR, USA). Gating strategies, a list of antibodies/fluorophores, and additional information are provided in Supporting information (Figure S2 and Table S1 ).
6
Phenotypic Analysis of Melanoma Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
FACS analysis was used for the detection of PD-1, CTLA-4, and PD-L1 (Becton Dickinson, Heidelberg, Germany) on the surface of melanoma cells in dependence of H-1PV infection. Furthermore, maturation markers such as CD80, CD83, and CD86 (Becton Dickinson, Heidelberg, Germany) were used to determine DC maturation, and Granzyme B (Becton Dickinson, Heidelberg, Germany) to represent T cell activity. FACS staining was performed as recommended by the manufacturer's data sheets. Shortly, cells were harvested and washed twice with Wash buffer (PBS+2.5% FBS). FACS antibodies were added in concentrations as recommended by the manufacturer's data sheets and incubated at room temperature. After 20 min cells were washed twice with wash buffer and measured immediately. For intracellular staining, cells were treated for 20 min with fixation/permeabilization buffer (Becton Dickinson, Heidelberg, Germany). After washing with Perm/Wash buffer (Becton Dickinson, Heidelberg, Germany) antibodies were added and incubated for 30 min at 4°C. Cells were washed twice and analyzed by FACS.
Fluorescence was measured with a minimum of 25,000 events per sample in a FACSCalibur (BD Bioscience, Heidelberg, Germany) according to the manufacturer's instructions. Data analyses were performed using Cell Quest Pro software (BD Bioscience).
Fluorescence was measured with a minimum of 25,000 events per sample in a FACSCalibur (BD Bioscience, Heidelberg, Germany) according to the manufacturer's instructions. Data analyses were performed using Cell Quest Pro software (BD Bioscience).
7
Multicolor FACS for Immune Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To determine the frequency of immune cells, multi‐color fluorescence‐activated cell sorting (FACS) analysis was performed using the following antibodies: anti‐CD3, CD4, CD8, CD14, CD15, CD25, CD80, CD45RA, HLA‐DR, FoxP3, CTLA‐4, PD‐1, PD‐L1, CCR4, CCD6, CXCR3, 4‐1BB, and OX40 (Becton Dickinson). Flow cytometry was done using the Becton Dickinson FACSAria II system.
8
Cell Surface Marker Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To measure the cell surface expression of CD47, HER2, EGFR, and CD20, cells were stained with conjugated primary antibodies (BD Biosciences) for 30 minutes at 4°C, washed, and resuspended in staining buffer. Macrophages were incubated with anti-CD16/32 (BD Biosciences) for 15 minutes at room temperature to block nonspecific binding and stained with CD14, CD68, CD80, CD163, CD206, PD-L1, and HLA-DR (BD Biosciences). Cells were acquired using a BD LSR X-20, and the data were analyzed using FlowJo software.
9
Investigating SRSF2 Function in Tumor-Infiltrating Lymphocyte Immune Response
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To investigate the functions of SRSF2 in the immune response of TILs against autologous tumor cell lines, TILs transfected with SRSF2 siRNA were co-cultured with autologous tumor cell lines. Then, flow cytometry was performed with an anti-CD8 antibody (Biolegend, 344704) and anti-IFN-γ antibody (BD Biosciences, 554552) on BD AccuriC6 (BD Biosciences). To examine the expression level of the immune checkpoints in the TILs and PBMCs, the frequency of immune checkpoint + and CD8 + TILs was assessed using flow cytometry on BD AccuriC6 (BD Biosciences). The antibodies against the immune checkpoints used for flow cytometry included PD-L1 (BD Biosciences, 561787), BTLA (BD Biosciences, 564802), CTLA-4 (BD Biosciences, 561717), LAG3 (BD Biosciences, 565617), and CD160 (BD Biosciences, 562351). All data were analyzed by FlowJo software.
10
Comprehensive Immunohistochemical Protein Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein expression was assessed by immunohistochemistry (IHC), as previously described.19 (link) Cancer cells on slides were scored by pathologists. For PD-1, the tumor-infiltrating lymphocytes (TILs) were scored. Protein was considered overexpressed, when the percent staining and intensity were above previously published and validated thresholds specific to each marker, for proteins where increased expression levels were of interest; when loss of protein or underexpression of protein was of interest, percent staining and intensity below previously published and validated levels were reported.19 (link) Antibodies used included: androgen receptor (AR), topoisomerases 1 and 2 (TOPO1, TOPO2A, Leica Biosystems), estrogen receptor (ER), progesterone receptor (PR), cMet, human epidermal growth factor receptor 2 (HER2; Ventana), cKIT, epidermal growth factor receptor (EGFR), phosophatase and tensin homolog (PTEN; Dako), O(6)-methylguanine-methyltransferase (MGMT), PGP, thymidylate synthase (TS; Invitrogen), transducin-like enhancer of split 3 (Santa Cruz), excision repair cross complimentary group 1 (ERCC1; Abcam), RRM1 (Proteintech), SPARC (monoclonal; R&D Systems and polyclonal; Exalpha) and tubulin beta-3 chain (Covance), programmed cell death protein 1 (PD-1), and programmed death-ligand 1 (PD-L1; BD Pharmingen and R&D Systems).
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!