Cd4 untouched kit

Manufactured by Miltenyi Biotec

The CD4 untouched kit is a laboratory product designed for the isolation of CD4+ T cells from a variety of sample types, including peripheral blood mononuclear cells (PBMCs). The kit utilizes a magnetic bead-based separation method to negatively select the CD4+ T cells, leaving them in an untouched state, suitable for further analysis or downstream applications.

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Lab products found in correlation

Moflo astrio, by Beckman Coulter (1 mentions) Anti cd90.1 microbeads, by Miltenyi Biotec (1 mentions) Flow count fluorosphere, by Beckman Coulter (1 mentions)

Spelling variants of this product

Untouched CD4+ T cell isolation kit (5 citations)

2 protocols using cd4 untouched kit

Isolation and Purification of Treg and Tconv Cells

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For analyses in FoxP3 proficient mice, splenic CD4 T cells from FoxP3-Thy1.1 hemizygous male mice were first enriched using CD4 untouched kit (Miltenyi), followed by secondary purification with anti-CD90.1 microbeads (Miltenyi) according to manufacturer’s instructions. Tconv (FoxP3-Thy1.1-Tcrb+CD4+CD45+CD19-CD8-CD11b-CD11c-) and Treg cells (FoxP3-Thy1.1+Tcrb+CD4+, CD45+CD19-CD8-CD11b-CD11c-) were sorted on a FACS-Aria to 96% and 94% purity, respectively. For experiments in FoxP3-deficient Tregs (Fig. 3), CD4⁺ T cells from spleen and peripheral lymph nodes from 6 week-old heterozygous females were first enriched as above by negative magnetic selection (Stem Cell), and Treg-like cells sorted as CD19-TCRβ+CD4+Thy1.1-GFP+ on a Moflo Astrios (Beckman Coulter).

Ramirez R.N., Chowdhary K., Leon J., Mathis D, & Benoist C. (2022). FoxP3 associates with enhancer-promoter loops to regulate Treg-specific gene expression. Science immunology, 7(67), eabj9836.

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Quantifying Epitope-Specific T Cell Responses

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In vaccination studies, CD4+ T cells from splenocytes of OT-2 or Marilyn mice were isolated by magnetic cell separation using the CD4 untouched kit (Miltenyi Biotec). Wildtype C57BL/6 mice were intraperitoneally (i.p.) injected with 4×10⁶ OT-2 specific or 2×10⁶ DBY-specific T-cells after in vitro expansion. Twenty-four hours later, mice were subcutaneously vaccinated with 10 µg of the indicated 30mer peptides using 50 µg Poly(I:C) or CpG with 50 µl IFA as adjuvant. Epitope-specific T cells were quantified from blood of vaccinated mice based on expression of CD4+/TCRvα2/β5+ (OT-2) or CD4+/TCRvβ6+/CD45.1+ (Marilyn). Cell numbers were quantified using flow count fluorospheres for normalization (Beckman Coulter). On day 7 after vaccination mice were euthanized and cells from spleen and lymph nodes were isolated for analysis.

Bernhardt A.L., Zeun J., Marecek M., Reimann H., Kretschmann S., Bausenwein J., van der Meijden E.D., Karg M.M., Haug T., Meintker L., Lutzny-Geier G., Mackensen A, & Kremer A.N. (2021). Influence of DM-sensitivity on immunogenicity of MHC class II restricted antigens. Journal for Immunotherapy of Cancer, 9(7), e002401.

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