Fitc anti cd4 antibody

Manufactured by BD

Sourced in United States

The FITC-anti-CD4 antibody is a fluorescently labeled monoclonal antibody that recognizes the CD4 surface antigen. It is commonly used in flow cytometry applications to identify and quantify CD4-positive cells in biological samples.

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Lab products found in correlation

Brefeldin a, by Thermo Fisher Scientific (2 mentions) Fixable viability stain 780, by BD (2 mentions) Ionomycin, by Thermo Fisher Scientific (1 mentions) Anti cd4 magnetic beads, by Miltenyi Biotec (1 mentions) Phorbol 12 myristate 13 acetate pma, by Thermo Fisher Scientific (1 mentions) Fix perm set, by BioLegend (1 mentions) 5 and 6 carboxyfluorescein diacetate succinimidyl ester cfse, by Thermo Fisher Scientific (1 mentions) Anti cd8 pe, by BD (1 mentions) Facscalibur, by BD (1 mentions) Mouse th1 th2 th17 cba kit, by BD (1 mentions) Cytofix cytoperm soln kit, by BD (1 mentions) Mouse beta actin antibody, by Proteintech (1 mentions) Leuko act cktl with golgiplug, by BD (1 mentions) Pe anti foxp3 antibody, by Thermo Fisher Scientific (1 mentions) Myeloperoxidase mpo kit, by Nanjing Jiancheng (1 mentions) Bca protein determination kit, by Beyotime (1 mentions) Trizol reagent, by Takara Bio (1 mentions) Sybr premix ex taq, by Takara Bio (1 mentions) Reactive oxygen species assay kit, by Beyotime (1 mentions) Transcription factor buffer set, by BD (1 mentions)

Close spelling variants of this product

Mouse anti-human CD4-FITC antibody FITC-labeled anti-mouse CD4 antibody FITC-conjugated anti-mouse CD4 antibody FITC-anti-mouse CD4 or PerCP-Cy5.5-anti-mouse CD8 antibody FITC Rat Anti–Mouse CD4 antibody FITC-conjugated anti-human CD4+ antibody FITC-conjugated anti-CD4 antibody FITC anti-mouse CD4 antibody Anti-CD4-FITC Anti-CD4 antibody

6 protocols using fitc anti cd4 antibody

CD4+ T Cell Activation and Proliferation Analysis

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Spleen CD4⁺ T cells derived from OT-II mice were purified using anti-CD4 magnetic beads (Miltenyi Biotec) as previously described (Jin et al., 2019b (link)). The purified CD4⁺ T cells had >90% purity. DCs (1 × 10⁵/well) and CD4⁺ T cells (1 × 10⁶/well) were cocultured for 72 h with OVA (1 mg/mL). To determine the cytokine production, cells were stimulated with 10 mg/mL Brefeldin A (eBioscience), 50 ng/mL phorbol 12-myristate 13-acetate (PMA) (eBioscience), and 750 ng/mL Ionomycin (eBioscience) for 6 h at 37°C. Cells were stained with FITC-anti-CD4 antibodies (BD Biosciences) for 35 min at 4°C. These cells were fixed, permeabilized using a FIX/PERM set (Biolegend) and blocked in 5% rat serum for 10 min at room temperature in the dark prior to intracellular staining with APC-conjugated mAbs to IFN-γ and PE-conjugated mAbs to IL-4 (Jin et al., 2020a ).
To determine CD4 + T-cell proliferation induced by DCs, CD4 + T cells (5 × 10⁵/well) were stained with 5-and 6-carboxyfluorescein diacetate succinimidyl ester (CFSE) (eBioscience) before co-culture with DCs. Samples were analyzed using a BD FACS Calibur Flow Cytometer and FlowJo software (Tree star Inc, Ashland, OR) (Jin et al., 2019b (link)).

Liu Y., Liu X., Yang L., Qiu Y., Pang J., Hu X., Dong Z., Liu Z, & Jin X. (2021). Adjuvanticity of β -Glucan for Vaccine Against Trichinella spiralis. Frontiers in Cell and Developmental Biology, 9, 701708.

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Evaluating CTPG's Anti-tumor Effects

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B16-F10 cells in log-phase growth were re-suspended in PBS at a density of 5×10⁶/ml. 5 × 10⁵B16-F10 cells were subcutaneously injected into the right flank of C57BL/6 mice. After 3 days, mice were randomly divided into 3 groups (12 mice/group). Control group was administered with 0.2 ml DMSO. CTPG-200 and CTPG-400 groups were subcutaneously administered with 200 or 400 mg/kg CTPG in 0.2 ml DMSO around tumor, respectively. Mice were treated every 2 days for up to 15 days. Tumor volume was measured using calipers and calculated according to the following formula: tumor volume (mm³) = (length×width²)/2. On day 31, the animals were sacrificed and spleens were isolated to analyze the percentages of CD8⁺T and CD4⁺ T cells. Splenocytes were stained with PE-anti-CD8 and FITC-anti-CD4 antibodies (BD Biosciences) for 15 min at RT. After washing with PBS, samples were analyzed by flow cytometry (BD FACSCalibur, USA).

Li J., Li J., Aipire A., Gao L., Huo S., Luo J, & Zhang F. (2016). Phenylethanoid Glycosides from Cistanche tubulosa Inhibits the Growth of B16-F10 Cells both in Vitro and in Vivo by Induction of Apoptosis via Mitochondria-dependent Pathway. Journal of Cancer, 7(13), 1877-1887.

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Pharmacological Targeting of Th17/Treg Imbalance in Murine Colitis

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Luteolin and DTT were obtained from Aladdin (China). 1,4-butanediol diacrylate (BDD) was purchased from TCI (Shanghai, China). Dextran sulfate sodium (DSS, 35000 Da) was purchased from MP Biomedical (USA). Mouse Th1/Th2/Th17 CBA Kit, Cytofix/CytopermSoln Kit, Leuko Act Cktl with GolgiPlug, Transcription Factor Buffer Set, Fixable Viability Stain 780, FITC-Anti-CD4 antibody, BV605-Anti-IL-17A antibody, APC-Anti–INF–γ antibody and PE-Anti-IL-4 antibody were purchased from BD Biosciences (San Diego, USA). PE-Anti-FOXP3 antibody was purchased from eBiosciences (San Diego, CA). Myeloperoxidase (MPO) kit, glutathione (GSH) kit and ROS kit were supplied by Nanjing Jiancheng Bioengineering Institute (Nanjing, China). The BCA protein determination kit and Reactive Oxygen Species Assay Kit were purchased from Beyotime Biotechnology (China). Mouse Beta Actin antibody, Mouse Occludin antibody and Mouse ZO-1 antibody for western blotting were purchased from Proteintech (China). SYBR Premix Ex Taq™, Trizol reagent and PrimeScript™ RT 131 Master Mix were obtained from TaKaRa Bio (China). All other reagents are commercially available and can be used directly. All the solvents used were of analytical grade and were procured from Sinopharm (China).

Tan C., Fan H., Ding J., Han C., Guan Y., Zhu F., Wu H., Liu Y., Zhang W., Hou X., Tan S, & Tang Q. (2022). ROS-responsive nanoparticles for oral delivery of luteolin and targeted therapy of ulcerative colitis by regulating pathological microenvironment. Materials Today Bio, 14, 100246.

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Isolation of CD4+ T Cells from Mouse Spleen

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Specific fluorescent antibody-labeled CD4⁺ T cells were separated from mouse spleen mononuclear cells by flow cytometry. C57BL/6 mice aged 10 weeks were euthanized by cervical dislocation to isolate splenocytes. Then, a single-cell suspension was prepared by using mechanical trituration method where the tissue was ground through a 300-mesh sieve. Subsequently, mouse spleen mononuclear cells were re-suspended using 50% and 70% Percoll separating media (GE Healthcare Life Sciences, USA). Density-gradient centrifugation was undertaken at 2500 rpm for 25 min following red blood cell lysis. The steps mentioned above were performed at a fast pace, at 4 °C or on ice. For cell surface staining, the antibodies (FITC-anti-CD4) were incubated with the single-cell suspension for at least 30 min at 4 °C. FITC-anti-CD4 antibody was purchased from BD Bioscience (USA). Cells were sorted with the flow cytometer FACS Aria II (Becton, Dickinson and Company).

Xu W., Wu Y., Wang L., Bai Y., Du Y., Li Y., Cao N., Zhao Y., Zhang Y, & Liu H. (2019). Autoantibody against β1-adrenoceptor promotes the differentiation of natural regulatory T cells from activated CD4+ T cells by up-regulating AMPK-mediated fatty acid oxidation. Cell Death & Disease, 10(3), 158.

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Quantification of Regulatory T Cells

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Hemolyzed peripheral blood samples or spleen cells were incubated with purified CD16/32 (cat#553142 BD Pharmingen (BD), San Jose, CA) to block nonspecific staining. Dead cells were defined as cells that were positive for Fixable Viability Stain 780 (cat# 565388 BD). Tregs that differentiated from naïve CD4+ cells were identified by a two-step staining process. First, cells were surface-stained with APC-anti-CD25 antibody (cat# 561048 BD) and FITC-anti-CD4 antibody (cat# 557667 BD). Subsequently, the cells were fixed and permeabilized with Mouse Foxp3 Buffer Set (cat#560409 BD), and then stained with PE anti-mouse Foxp3 antibody (cat# 560414 BD). Cell samples were analyzed on a BD FACS VERSE flow-cytometer (BD). Gating strategy was shown in Supplemental Fig. S2.

Mei T., Noguchi H., Kuraji R., Kubo S., Sato Y., Kaku K., Okabe Y., Onishi H, & Nakamura M. (2023). Effects of periodontal pathogen-induced intestinal dysbiosis on transplant immunity in an allogenic skin graft model. Scientific Reports, 13, 544.

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Th2 Polarization of Naive T Cells

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Splenic naive T cells from DO11.10 mice were purified using a MagCellect kit from R&D Systems according to the manufacturer's instructions. Th2 cell polarization were performed as described elsewhere. 32 , 33 Briefly, 1 × 10 6 naive T cells were co-cultured with 0•5 × 10 6 sorted and hemin or hemin plus SnPP pre-treated BMBs in 48-well plates in the presence of 20 U/ml IL-2 (R&D Systems), 10 ng/ml IL-3, 100 μg/ml Dinitrophenyl (DNP)-OVA (Biosearch Technologies, Novato, CA), and 10 μg/ml anti-DNP IgE (Sigma-Aldrich). As control, 1 × 10 6 naive T cells alone were used. Fresh complete RPMI-1640 medium with 10 U/ml IL-2 was added at day 3. After 5 days of culture, cells were stimulated by 20 ng/ml PMA (Sigma-Aldrich) and 1 μg/ml ionomycin (Sigma-Aldrich) for a further 6 hr. Two hours before collection, Brefeldin A (eBioscience) was added into the culture medium at the ratio of 1 : 1000. The collected cells were surface stained with FITC-anti-CD4 antibody (BD Pharmingen). After washing, fixing and permeabilizing, cells were intracellularly stained with PE-anti-IL-4 and APC-anti-interferon-γ antibodies (eBioscience), and examined with flow cytometry to detect the Th1/Th2 subsets.

Zhong W., Di C., Lv J., Zhang Y., Lin X., Yuan Y., Lv J, & Xia Z. (2016). Heme oxygenase-1 inhibits basophil maturation and activation but promotes its apoptosis in T helper type 2-mediated allergic airway inflammation. Immunology, 147(3).

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