Anti cd28 antibodies

Manufactured by BioXCell

Sourced in United States

Anti-CD28 antibodies are immunological reagents that bind to the CD28 receptor on T cells. CD28 is a co-stimulatory molecule that plays a crucial role in T cell activation and proliferation. These antibodies can be used in research applications to study T cell biology and immune responses.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

Anti-CD3 and anti-CD28 antibodies Anti-CD28 antibodies (clone 37.51; Anti-CD28

5 protocols using anti cd28 antibodies

T Cell Activation Signaling Assay

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

MTT (1-(4,5-Dimethylthiazol-2-yl)-3,5-diphenylformazan) powder, CMFDA (5-chloromethylfluorescein diacetate) and CMRA (5-(((4-chloromethyl)benzoyl)amino) tetramethylrhodamine) cell staining dyes, was obtained from Sigma Chemical Co. (St. Louis, MO, USA). Anti-CD3 antibodies and anti-CD28 antibodies were purchased from Bioxcell (West Lebanon, NH, USA). Human IL-2 DuoSet^® ELISA kit was obtained from R&D Systems (Minneapolis, MN, USA). Staphylococcal enterotoxin E (SEE) was purchased from Toxin Technology (Sarasota, FL, USA). NE-PER Nuclear and Cytoplasmic Extraction Reagents Kit and ECL Western blotting detection reagents were obtained from Thermo Fisher Scientific (Waltham, MA, USA). Anti-CD40L antibodies conjugated with APC was purchased from eBiosciences (San Diego, CA, USA). Anti-CD40L neutralizing antibodies were obtained from InvivoGen (San Diego, CA, USA). Anti-CD40L for western blot and anti-β-actin antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Anti-p65, anti-PARP, anti-IκBα, anti-phosphorylated IκBα (S32), anti-phosphorylated ERK (T202/Y204), anti-ERK, anti-phosphorylated p38 (T180/Y182), anti-p38, anti-phosphorylated JNK (T183/Y185), anti-JNK, anti-phosphorylated c-Jun (S73) and anti-c-Jun antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA).

Lee H.S, & Jeong G.S. (2020). Chrysophanol Mitigates T Cell Activation by Regulating the Expression of CD40 Ligand in Activated T Cells. International Journal of Molecular Sciences, 21(17), 6122.

+ Open protocol

+ Expand

Murine T Cell Isolation and Culture

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

The spleens of 6‐to 8‐week‐old C1QBP^+/− and C1QBP^+/+ C57/BL6 mice were harvested, gently ground in MACS buffer (PBS + 0.5% BSA + 2 mM EDTA) and then passed through a sterile 40‐µm nylon filter (BD Falcon). Red blood cells were lysed with RBC Lysing Buffer (BioLegend). The murine T cells were isolated by Pan T magnetic Microbeads (Miltenyi Biotec) from splenocytes obtained from C1QBP^+/+ and C1QBP^+/− mice. CD4⁺ or CD8⁺ T cells were sorted on a FACSAria III Cell Sorter (BD Biosciences) and planted in 24‐well plates coated with 2.5 μg/mL anti–CD3ε (145‐2C11, Bio‐X‐Cell) and 1 μg/mL anti–CD28 antibodies (37.51 Bio‐X‐Cell). All the above cells were cultured in RPMI‐1640 medium (Corning) supplemented with 10% FBS, 4 mM l‐glutamine, 1% penicillin/streptomycin, 2 ng/mL IL‐2, 10 ng/mL IL‐7, 5 ng/mL IL‐15, and 75 μM β‐mercaptoethanol and incubated at 37°C in a 5% CO₂ humidified atmosphere.

Tian H., Wang G., Wang Q., Zhang B., Jiang G., Li H., Chai D., Fang L., Wang M, & Zheng J. (2022). Complement C1q binding protein regulates T cells’ mitochondrial fitness to affect their survival, proliferation, and anti–tumor immune function. Cancer Science, 113(3), 875-890.

+ Open protocol

+ Expand

Efficient in vitro differentiation of murine T helper cell subsets

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

Mouse CD4⁺ T cells were isolated from spleens using the MagniSort Mouse CD4⁺ T cell enrichment kit (Invitrogen, 8804-6821-74) following the manufacturer’s instructions. Cells were maintained in complete RPMI1640 medium (Corning, 10-040-CV) containing 10% FBS, 2 mM l-glutamine, 50 U/ml penicillin-streptomycin, and 5.5 mM β-mercaptoethanol. For T_H2 cell differentiation, naïve CD4⁺ T cells isolated from WT and Orai1^fl/flCd4Cre mice were stimulated with anti-CD3 (2.5 ng/ml; clone 2C11) and anti-CD28 antibodies (1 μg/ml; clone 37.51) (both Bio X Cell) on flat-bottom plates coated with rabbit anti-hamster IgG (25 μg/ml; MP Biomedicals, catalog no. MP0855398) in the presence of IL-4 (50 ng/ml; PeproTech) and anti-IFN-γ (5 μg/ml; eBioscience, clone 11B11) for 2 days. For T_H1 differentiation, CD4⁺ T cells were stimulated with anti-CD3 (1 μg/ml) and anti-CD28 (1 μg/ml) antibodies on IgG-coated plates as described above in the presence of IL-12 (20 ng/ml; PeproTech) and anti-IL-4 (5 μg/ml; eBioscience, clone 11B11). On day 2, cells were detached and expanded in RPMI1640 medium supplemented or not with T_H1 or T_H2 cytokines as described above and hIL-2 (20 U/ml; PeproTech, 200-02). Cells were analyzed on days 3 to 5.

Wang Y.H., Noyer L., Kahlfuss S., Raphael D., Tao A.Y., Kaufmann U., Zhu J., Mitchell-Flack M., Sidhu I., Zhou F., Vaeth M., Thomas P.G., Saunders S.P., Stauderman K., Curotto de Lafaille M.A, & Feske S. (2022). Distinct roles of ORAI1 in T cell–mediated allergic airway inflammation and immunity to influenza A virus infection. Science Advances, 8(40), eabn6552.

+ Open protocol

+ Expand

Naïve CD4+ T Cell Differentiation

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

Naïve CD4⁺ T cells (CD25⁻Foxp3-GFP⁻CD4⁺CD8⁻TCRβ⁺CD62L⁺CD44⁻) cells were sorted from pooled spleens and lymph nodes after CD4⁺ cell enrichment using the Dynabeads™ FlowComp™ Mouse CD4 Kit according to the manufacturer’s instructions. Tissue culture treated 96-well flat-bottom plates (USA Scientific) were coated with 1 μg/mL anti-mouse CD3 and 1 μg/mL anti-mouse CD28 antibodies (BioXCell) in 200 μL 1x PBS at 37°C for 2 hours and then washed once with 1x PBS. Cells were cultured on these plates for 2–4 days with 1 ng/ml recombinant human TGF-β and varying concentrations of recombinant human IL-2 before staining and analysis. To assay the stability of Foxp3 expression, naïve CD4⁺ T cells were cultured in Treg cell induction conditions with or without 0.25 mM ascorbic acid-2-phosphate (Sigma) for 4 days. Foxp3-GFP⁺ cells were then sorted and cultured on new plates coated with or without 1 μg/mL anti-CD3 and 1 μg/mL anti-CD28 antibodies (BioXCell) in the presence of 100 U/mL recombinant IL-2 for 1–4 days. At the end of culture, cells were harvested and stained with a viability dye (BioLegend) followed by cell fixation and Foxp3 staining.

Dikiy S., Li J., Bai L., Jiang M., Janke L., Zong X., Hao X., Hoyos B., Wang Z.M., Xu B., Fan Y., Rudensky A.Y, & Feng Y. (2021). A distal Foxp3 enhancer enables interleukin-2 dependent thymic Treg cell lineage commitment for robust immune tolerance. Immunity, 54(5), 931-946.e11.

+ Open protocol

+ Expand

Isolation and Proliferation of Naïve T Cells

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

Naïve T cells were prepared using a T cell isolation kit (STEMCELL, # 19851) with > 97% purity from the spleens of BALB/c mice. The purified cells were labeled with CFSE dye (Invitrogen, # C34554) for the proliferation assay then cultured in the complete medium supplemented with HEPES, β-mercaptoethanol, and anti-CD28 antibodies (2.5 μg/ml; Bio X Cell, # BE0015-1) in an anti-CD3 antibody pre-coated plate (1 μg/ml; Bio X Cell, # BE0001-1).

Hawthorne G., Irgens L.M, & Lie R.T. (2000). Outcome of pregnancy in diabetic women in northeast England and in Norway, 1994-7. BMJ : British Medical Journal, 321(7263), 730-731.

+ 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!