Mf 14

Manufactured by BioLegend

Sourced in China, United States

The MF-14 is a motorized microplate fixture designed for use with BioLegend's microplate readers. It provides automated, precise, and reproducible positioning of microplates during read operations.

Automatically generated - may contain errors

Lab products found in correlation

6 protocols using mf 14

Murine Immune Cell Profiling

Check if the same lab product or an alternative is used in the 5 most similar protocols

Antibodies against CD45 (104), CD3 (17A2), NK1.1 (PK136), CD19 (6D5), CD69 (H1.2F3), CD4 (RM4-4), CD8 (53-6.7), Ki-67 (11F6), CD146 (ME-9F1), Ly6G (1A8), CD11b (M1/70), F4/80 (BM8), CD11c (N418), MHC-II (M5/114.15.2), CD86 (GL-1), CD80 (16-10A1), PD-1 (RMP1-14), and FoxP3 (MF-14) were purchased from BioLegend. The fixable viability dye eFluor506 was purchased from eBioscience. Liver cell suspensions were subjected to surface staining with fluorescently labeled antibodies according to the manufacturer’s instructions. Cell viability was assessed using the fixable viability dye eFluor506 (eBioscience). Subsequently, cells were permeabilized using the Transcription Factor Buffer Set (Biolegend) and stained for Ki-67, FoxP3. Cells were analyzed on a CytoFLEX flow cytometer (Beckman Coulter, United States). Flow cytometry data were analyzed using FlowJo software (FlowJo, Ashland, OR, United States).

Chen L., Zheng H., Yu X., Liu L., Li H., Zhu H., Zhang Z., Lei P, & Shen G. (2020). Tumor-Secreted GRP78 Promotes the Establishment of a Pre-metastatic Niche in the Liver Microenvironment. Frontiers in Immunology, 11, 584458.

+ Open protocol

+ Expand

Comprehensive Immune Cell Profiling in Tumor and Lymph Nodes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Approximately 1*10⁶ cells from DLN or 5*10⁶ cells from tumors were stained for 30 min at room temperature with Zombie Red Fixable Viability Kit (Biolegend) and afterwards incubated on ice for 20 min with TruStain fcX (anti-mouse CD16/32, Biolegend). Surface staining was performed with APC-coupled AH1 tetramers and fluorochrome-conjugated antibodies against Thy1.2 (clone 53-2.1), CD8 (53-6.7), CD4 (GK1.5), CD44 (IM7), CD62L (MEL-14), PD-1 (29F.1A12), TIM-3 (RMT3-23) and LAG-3 (C9B7W), which were all purchased from Biolegend. Cells were stained in PBS containing 0.5% bovine serum albumin (BSA) and 2 mM EDTA for 1 h at 4ºC. For the staining of intracellular markers, cells were fixed and permeabilized using the eBioscience Foxp3 / Transcription Factor Staining Buffer Set according to the protocol of the manufacturer. Cells were incubated with fluorochrome-conjugated antibodies against Ki-67 (16A8, Biolegend) and Foxp3 (MF-14, Biolegend) in PBS containing 0.5% BSA and 2 mM EDTA for 30 min at room temperature. Cells were analyzed on a CytoFLEX cytometer (Beckman Coulter) and data were processed using FlowJo (v.10, Tree Star).

Probst P., Stringhini M., Ritz D., Fugmann T, & Neri D. (2018). Antibody-based delivery of TNF to the tumor neo-vasculature potentiates the therapeutic activity of a peptide anti-cancer vaccine. Clinical cancer research : an official journal of the American Association for Cancer Research, 25(2), 698-709.

+ Open protocol

+ Expand

Murine Immune Cell Phenotyping

Check if the same lab product or an alternative is used in the 5 most similar protocols

Fluorochrome-conjugated antibodies specific for mouse CD4 (GK1.5), CD8a (53–6.7), CD5 (53–7.3, isotype: rat IgG2a, κ), IFN-γ (XMG1.2), CD45 (30-F11, isotype: rat IgG2b, κ), CD326 (EpCAM, G8.8, isotype: rat IgG2α, κ), I-Ab (AF6-120.1), CD11c (N418), and FoxP3 (MF-14), and human CD3 (HIT3a), CD8a (HIT8a), CD4 (OKT4), CD45RA (Hl100), CD45RO (UCHL1), and CD62L (DREG-56) were obtained from BioLegend. Mouse CD3ε (145-2C11), CD69 (H1.2F3, isotype: Armenian hamster IgG), Ly51 (6C3, isotype: rat IgG2a, κ), and HLA-DR (L243) specific antibodies were purchased from BD. The TCR Vβ repertoire kit (IOTest Beta Mark) was purchased from Beckman Coulter. UEA I lectin was obtained from GeneTex. Thymus, spleen, and LNs from 1–2-mo-old C57BL/6, ABabDII and ABabDR4 mice were isolated. Cells were obtained by mashing the organs through a 0.45-µm cell strainer. Isolation of thymic DCs and epithelial cells was performed as published (Xing and Hogquist, 2014 (link)). In brief, thymic lobes were digested in enzyme solution (RPMI-1640 medium with 0.05% Liberase TH and 100 U/ml of DNase I) at 37°C for 20 min. Single cells were then stained with antibodies specified in the respective figure legends and analyzed by flow cytometry (FACSCanto II; BD). FoxP3 staining was performed with True-Nuclear transcription factor buffer set from BioLegend.

Chen X., Poncette L, & Blankenstein T. (2017). Human TCR-MHC coevolution after divergence from mice includes increased nontemplate-encoded CDR3 diversity. The Journal of Experimental Medicine, 214(11), 3417-3433.

+ Open protocol

+ Expand

Phenotyping Immune Cells by Flow Cytometry

Check if the same lab product or an alternative is used in the 5 most similar protocols

Recombinant mouse Fms-like tyrosine kinase 3 (Flt3) ligand was purchased from Peprotech (Suzhou, China). Fluorescein-conjugated monoclonal antibodies against CD45 (30-F11), CD11c (N418), CD11b (M1/70), adhesion G protein-coupled receptor E1 (F4/80, BM8), Gr-1, programmed cell death ligand 1 (PD-L1, 10

.9G2), CD3 (17A2), CD4 (GK1.5), CD8 (53

–

6.7), IFNγ (XMG1.2), and FOXP3 (MF-14">MF-14); isotype control monoclonal antibodies, and carboxyfluorescein diacetate succinimidyl ester (CFSE) were purchased from Biolegend (Beijing, China); leukocyte activation cocktail (LAC) was purchased from BD Pharmingen (Shanghai, China). Microbead-conjugated monoclonal antibodies against CD8 or CD11c were purchased from Miltenyi Biotec (Shanghai, China). The iTAg Tetramer/PE-H-2Kb OVA (SIINFEKL) detection kit was purchased from MBL (Shanghai, China). Ovalbumin peptide SIINFEKL (OVA257-264), collagenase type IV, hyaluronidase, and deoxyribonuclease were purchased from Sigma-Aldrich (Shanghai, China). Dextran sulfate sodium (DSS; molecular mass 36,000–50,000 Da) was purchased from MP Biomedicals (Beijing, China). Monoclonal antibodies against human CD8 (EP1150Y) and HDAC9 (EPR5223) were purchased from Abcam (Shanghai, China).

Ning Y., Ding J., Sun X., Xie Y., Su M., Ma C., Pan J., Chen J., Jiang H, & Qi C. (2020). HDAC9 deficiency promotes tumor progression by decreasing the CD8+ dendritic cell infiltration of the tumor microenvironment. Journal for Immunotherapy of Cancer, 8(1), e000529.

+ Open protocol

+ Expand

Multi-Marker Immune Profiling of T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Anti-mouse: α-CD45 (30-F11, BD, 4 μg ml⁻¹), α-CD3 (145-2C11, Biolegend, 5 μg ml⁻¹), α-CD4 (GK1.5, Biolegend, 5 μg ml⁻¹), α-CD8 (53-6.7, BD, 5 μg ml⁻¹), α-CTLA-4 (1B8, abcam, 30 μg ml⁻¹), α-TIM3 (B8.2C12, Biolegend, 2.5 μg ml⁻¹), α-OX40 (OX-86, Biolegend, 10 μg ml⁻¹), α-CD62L (MEL-14, BD, 5 μg ml⁻¹), α-CD44 (IM7, Biolegend, 2.5 μg ml⁻¹), α-LAG-3 (C9B7W, Biolegend, 5 μg ml⁻¹), α-IFN-γ (XMG1.2, Biolegend, 2.5 μg ml⁻¹), α-TNF-α (MP6-XT22, Biolegend, 2.5 μg ml⁻¹), α-Granzyme B (NGZB, ThermoFisher, 1.25 μg ml⁻¹), α-FOXP3 (MF-14, Biolegend, 10 μg ml⁻¹), α-Ki67 (SolA15, ThermoFisher, 0.6 μg ml⁻¹). Anti-human: α-CD4 (A161A1, Biolegend, 2.5 μg ml⁻¹), α-T-bet (4B10, Biolegend, 2.5 μg ml⁻¹), α-Phospho-Akt (Ser473) (D9E, Cell Signaling, 0.5 μg ml⁻¹), α-Phospho-Akt (Thr308) (D25E6, Cell Signaling, 0.5 μg ml⁻¹), α-Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E, Cell Signaling, 0.5 μg ml⁻¹).

Ho W.S., Wang H., Maggio D., Kovach J.S., Zhang Q., Song Q., Marincola F.M., Heiss J.D., Gilbert M.R., Lu R, & Zhuang Z. (2018). Pharmacologic inhibition of protein phosphatase-2A achieves durable immune-mediated antitumor activity when combined with PD-1 blockade. Nature Communications, 9, 2126.

+ Open protocol

+ Expand

Isolation and Analysis of Mesenteric Lymph Nodes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mesenteric lymph nodes (mLN) of DSS treated animals were isolated, passed through a 100-µm cell strainer and red blood cells were lysed with the RBC lysis buffer (Biolegend, San Diego, CA, USA). To differentiate between live and dead cells, cells were stained for 15 min at room temperature with the Zombie UV Fixable Viability Kit (Biolegend, San Diego, CA, USA) as per the company's recommendation. Cell surface staining was performed for 30 min at 4°C with fluorochrome-conjugated monoclonal antibodies to mouse antigens purchased from Biolegend (San Diego, CA, USA): CD45 (30-F11), CD11c (N418), PDCA-1 (927), CD3 (17A2), CD4 (GK1.5), CD25 (PC61). Following staining cells were washed and fixed in 4% PF.
For intracellular staining, cells first were stained for cell surface markers, washed, then fixed, permeabilized and stained for intracellular transcription factor FoxP3 (MF-14; Biolegend, San Diego, CA, USA) for 30 min at RT. Samples were analyzed by the FACSAria cell sorter (BD Biosciences, Franklin Lakes, NJ, USA), and the acquired data were analyzed using FlowJo software (version 10.6.1) (Ashland, OR, USA).

Halasi M., Grinstein M., Adini A, & Adini I. (2022). Fibromodulin Ablation Exacerbates the Severity of Acute Colitis. Journal of Inflammation Research, 15, 4515-4526.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!