Anti cd3

Manufactured by Jackson ImmunoResearch

Anti-CD3 is a primary antibody that specifically binds to the CD3 complex on the surface of T cells. This binding is a critical step in the activation and function of T cells, which play a central role in cell-mediated immune responses.

Automatically generated - may contain errors

Lab products found in correlation

Anti cd28, by Jackson ImmunoResearch (3 mentions) F ab 2 fragment of goat anti mouse igm, by Jackson ImmunoResearch (1 mentions) Ril 4, by R&D Systems (1 mentions) Anti cd28, by Thermo Fisher Scientific (1 mentions) Ki 67 rm 9106 s1, by Thermo Fisher Scientific (1 mentions) Facs antibodies, by Thermo Fisher Scientific (1 mentions) Permeabilization buffer, by Thermo Fisher Scientific (1 mentions) Fixation buffer, by Thermo Fisher Scientific (1 mentions) Anti cd3 cd28, by Merck Group (1 mentions) Brefeldin a, by Merck Group (1 mentions) Lipopolysaccharides (lps), by BioLegend (1 mentions) Anti cd40, by BioLegend (1 mentions)

5 protocols using anti cd3

Activation and Proliferation Assays for T and B Cells

Cited 2 times

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

CD3⁺ (T cells) or B220⁺ (B cells) cells isolated (Miltenyi Biotec, Inc, per manufacturers’ directions) from WT or S5A splenocytes were incubated overnight with anti-CD3 (1 μg/ml; 145–2C11, eBioscience)/anti-CD28 (1 μg/ml; 37.51, eBioscience) (T cells) or plate-bound anti-IgM 10 μg/ml; F(ab’)2 fragment of goat anti-mouse IgM, Jackson ImmunoResearch, West Grove, PA) (B cells). Upregulation of CD69 and CD86 was assessed by flow cytometry (6 (link), 24 (link)).
For proliferation assays, CD3⁺ (T cells) or B220⁺ cells (B cells) isolated from WT or S5A splenocytes using negative selection (Miltenyi Biotec Inc.) and were labeled with CTV (Invitrogen). Cells were incubated for 72 h in the absence or presence of anti-CD3/anti-CD28 (T cells) or of soluble anti-IgM stimulation (10 μg/ml; F(ab’)2 fragment of goat anti-mouse IgM, Jackson ImmunoResearch, West Grove, PA), and rIL-4 (10 ng/ml, R&D Systems) (B cells). Cells were analyzed for CTV dilution by flow cytometry.

Anaya E.P., Lin X., Todd E.M., Szasz T.P, & Morley S.C. (2021). Novel mouse model reveals that serine phosphorylation of L-plastin is essential for effective splenic clearance of pneumococcus. Journal of immunology (Baltimore, Md. : 1950), 206(9), 2135-2145.

+ Open protocol

+ Expand

Antibody-Based Tumor and Lineage Analysis

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

Antibodies used for tumor analysis include F4/80 (ab6640, Abcam, Cambridge, MA, USA) [99 (link)], EphA2 (C-20, SC-924, Santa Cruz Biotech, Dallas, TX, USA), Ki-67 (RM-9106-S1, Thermo Fisher Scientific, Waltham, MA, USA), SREBP1 (H-160), SREBP2 (H-164), FASN (H-300), and CD31 (CM303A, Biocare Medical, Concord, CA, USA). Antibodies used for lineage analysis were: anti-B220 (RA3-6B2), anti-CD19 (eBio1D3), anti-IgM µ-chain (Jackson Labs), anti-CD43 (S7) for B cells, anti-CD3 (145-2C11), anti-CD4 (GK1.5), anti-CD8a (53-6.7) for T cells, anti-CD41 (MWReg30), anti-CD71 (C2), anti-Ter119 (TER-119), and anti-F4/80 (BM8), anti-Mac1 (M1/70), and anti-Gr1 (RB6-8C5) for myeloid cells. Antibodies used for HSC and progenitor analysis were: Lineage (biotin-conjugated anti-Gr-1 (RB6-8C5), -Mac1 (M1/70), -B220 (RA3-6B2), -CD19 (eBio1D3), -Ter110 (TER-119), -CD5 (53-7.3), -CD4 (GK1.5), -CD8 (53-6.7)), APC-Cy7-c-Kit (2B8), PerCP-cy5.5-Sca1 (E13-161.7 or D7, 1:1000 dilution), FITC-CD48 (HM48-1), PE-Cy7-CD150 (TC15-12F12.2), APC-CD34 (RAM34) and PE–Flk2 (A2F10.1). All antibodies were used at 1:200 dilution unless otherwise noted. FACS antibodies were purchased from eBioscience (San Diego, CA, USA), BD Biosciences (San Jose, CA, USA) or BioLegend (San Diego, CA, USA).

Jiao X., Tian L., Zhang Z., Balcerek J., Kossenkov A.V., Casimiro M.C., Wang C., Liu Y., Ertel A., Soccio R.E., Chen E.R., Liu Q., Ashton A.W., Tong W, & Pestell R.G. (2021). Pparγ1 Facilitates ErbB2-Mammary Adenocarcinoma in Mice. Cancers, 13(9), 2171.

+ Open protocol

+ Expand

Spermidine Enhances T-cell Activation

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

For intracellular staining, PBMCs were stimulated in R10 with anti-CD3 (1 μg/ml, Jackson Immuno Research) and anti-CD28 (1 μg/mL, Jackson Immuno Research) with or without 10 μM spermidine for 4 days. On day 4, cells were re-stimulated with the same concentrations of anti-CD3/CD28 for 6 hr at 37°C in the presence of 1 μg/mL brefeldin-A (Sigma-Aldrich). As a control, cells were left unstimulated. Following surface marker staining as described above, cells were fixed with 100 μL Fixation buffer (eBioscience) for 20 min at RT in the dark. Next, cells were permeabilized with 100 μL of 1x Permeabilization buffer (eBioscience) for 15 min in the dark at RT. Then, cells were resuspended in the intracellular antibody mix (anti-IFN-γ, anti-Granzyme B, anti-Perforin) and incubated for 30 min in the dark at RT. After being washed twice with Permeabilization buffer the cells were resuspended in 200 μL of FACS buffer for analysis.

Alsaleh G., Panse I., Swadling L., Zhang H., Richter F.C., Meyer A., Lord J., Barnes E., Klenerman P., Green C, & Simon A.K. (2020). Autophagy in T cells from aged donors is maintained by spermidine and correlates with function and vaccine responses. eLife, 9, e57950.

+ Open protocol

+ Expand

Analyzing Tfh-like Cell Signaling

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

For Tfh-like cultures, on d2 or d3 postactivation (to allow expression of VHL and HIF1a which are not expressed in naïve cells), cells were stained with fixable viability dye, fixed in 4% paraformaldehyde, permeabilized in methanol at −20C for at least 1 h, washed with PBS once, then stained in FACS buffer with phospho-protein antibodies.
SgRNA or control transduced T cells were cultured in fresh complete RPMI 10% FCS with 10 U/ml hIL-2 daily, from day 3 to day 5 postactivation. Cells were rested in RPMI 1% FCS for 2 h, replated in fresh RPMI 1% FCS and restimulated by adding 1 ug/ml anti-CD3, 3 ug/ml anti-CD28, and 5 ug/ml goat anti-hamster (Jackson Immunoresearch) for 4 h. Cells were stained with viability dye, fixed, permeabilized, and stained as described above.

Huang B., Phelan J.D., Preite S., Gomez-Rodriguez J., Johansen K.H., Shibata H., Shaffer AL I.I.I., Xu Q., Jeffrey B., Kirby M., Anderson S., Yang Y., Gossa S., McGavern D.B., Staudt L.M, & Schwartzberg P.L. (2022). In vivo CRISPR screens reveal a HIF-1α-mTOR-network regulates T follicular helper versus Th1 cells. Nature Communications, 13, 805.

+ Open protocol

+ Expand

In vitro B and T Cell Assays

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

In vitro culture assays were performed as previously described(35 (link)). Briefly, 5x104 total CD19+CD4− B cells or GC B cells (CD19+CD4−Fas+GL-7+) 3x104 TFH cells and/or 1.5x104 TFR cells were plated in 96-well plates along with 25 ng LPS, 5 ug anti-CD40 (BioLegend) or 2ug/ml anti-CD3 and 5ug/ml anti-IgM (Jackson Immunoresearch). Polarizing cytokines for T cells and B cells class-switch recombination are described in specific experiments. Cultures were harvested after 3-6 days as specified in specific experiments. Recombinant tetrameric mouse Fgl2-His (R&D Systems) was added into the culture as described. For analysis, B cells were gated as CD19+CD4− cells while specific isotype staining was done by intracellular staining described earlier, TFH cells were gated as CD4+CD19−FoxP3− cells, and TFR cells were gated as CD4+ CD19−FoxP3+ cells.

Sungnak W., Wagner A., Kowalczyk M.S., Bod L., Kye Y.C., Sage P.T., Sharpe A.H., Sobel R.A., Quintana F.J., Rozenblatt-Rosen O., Regev A., Wang C., Yosef N, & Kuchroo V.K. (2020). TFR-derived Fibrinogen-like Protein 2 regulates production of autoantibodies and induction of systemic autoimmunity. Journal of immunology (Baltimore, Md. : 1950), 205(12), 3247-3262.

+ 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!