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Biotin protein l

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Biotin-Protein L is a conjugate of biotin and Protein L, a bacterial protein that binds to immunoglobulins. It is a versatile tool used in various immunoassays and protein purification applications.

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Lab products found in correlation

Plenti6.4 r4r2 v5 dest, by Thermo Fisher Scientific (2 mentions) Apc strept, by GenScript (2 mentions) Ficoll paque premium, by GE Healthcare (1 mentions) Anti cd3 fitc, by BioLegend (1 mentions) Fetal bovine serum (fbs), by Corning (1 mentions) Interleukin 2 (il 2), by Thermo Fisher Scientific (1 mentions) Il 15, by Thermo Fisher Scientific (1 mentions) Human cd3 microbeads, by Miltenyi Biotec (1 mentions) X vivo 15 media, by Lonza (1 mentions) Facsaria 2, by BD (1 mentions) Facscalibur, by BD (1 mentions) Anti cd4 buv395, by BD (1 mentions) Anti cd3 pe cy7, by BioLegend (1 mentions) Anti cd8 bv510, by BioLegend (1 mentions) Anti cd45ro percp cy5, by BioLegend (1 mentions) Anti cd45ra bv421, by BioLegend (1 mentions) Anti cd25 bv421, by BioLegend (1 mentions)

Spelling variants of this product

L-Biotin (2 citations) Biotin-labeled protein L (2 citations)

4 protocols using biotin protein l

1

Characterization of CAR-T Cell Therapy

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The University of Colorado Institutional Review Board approved the tissue bank protocol (IRB Protocol 06–0720), and all subjects gave informed consent in accordance with the Declaration of Helsinki. Healthy donor derived T cells were obtained from fresh peripheral blood mononuclear cells (PBMCs) using Ficoll-Paque PREMIUM (GE Healthcare, Chicago, IL). Patient derived T cells were obtained from cryopreserved peripheral blood from our MDS patient tissue bank. T cells were selected from the tissue samples using human CD3 MicroBeads (Miltenyi Biotec, Bergisch Gladbach, Germany). Purified CD3+ cells were cultured in X-VIVO™15 media (Lonza Group, Basel, Switzerland) with 10% heat-inactivated fetal bovine serum (Corning Inc., Corning, NY) and 50 unit/ml IL-2 (PeproTech, NJ) and 0.5ng/ml IL-15 (PeproTech). Cultured T cells were stimulated by CD3/CD28 Dynabeads (Gibco™ Biosciences) then transduced with CD123 CAR or left untransduced (mock). After 6~8 days, CD3/CD28 Dynabeads were removed from T cells, and CD123 CAR expression in transduced T cells was determined by flow with anti-CD3-FITC (BioLegend, CA), anti-human EGFRt (BioLegend), biotin-Protein L (GenScript, China), Straptevidin-PE (BD Biosciences, CA), and Live/Dead-APC/Cy7 viability dye (Invitrogen™ Thermo Fisher Scientific, MA).
Stevens B.M., Zhang W., Pollyea D.A., Winters A., Gutman J., Smith C., Budde E., Forman S.J., Jordan C.T, & Purev E. (2019). CD123 CAR T Cells for the Treatment of Myelodysplastic Syndrome. Experimental hematology, 74, 52-63.e3.
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2

T Cell Immunophenotyping by Flow Cytometry

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We applied BD FACSCalibur and BD FACSAria II instruments for FCM detection. BD FACSDiva software and FlowJo software were used for analysis. FCM detection was performed according to the schedule shown in Figure 2C. CD3, CD4, CD8, and CAR expression were evaluated on days 6, 10, and 14, and memory T subsets were evaluated on days 10 and 14. We used His-tagged recombinant CEACAM5 (CEA-His; Sino Biological Inc., 11077-H08H-50) and biotin-protein L (GenScript, M00097) for specific detection of CAR expression. The following antibodies were used: anti-CD3-PE-Cy7 (BioLegend, 300420), anti-CD4-BUV395 (BD, 564724), anti-CD8-BV510 (BioLegend, 344732), anti-CD25-BV421 (BioLegend, 302630), anti-CD45RA-BV421 (BioLegend, 304130), anti-CD45RO-PerCP-Cy5.5 (BioLegend, 304222), AF-647-conjugated IgG fraction of mouse monoclonal anti-biotin (Jackson, 200-602-211) and anti-CD197-PE (BioLegend, 353204).
Zhang C., Wang L., Zhang Q., Shen J., Huang X., Wang M., Huang Y., Chen J., Xu Y., Zhao W., Qi Y., Li Y., Ou Y., Yang Z, & Qian C. (2023). Screening and characterization of the scFv for chimeric antigen receptor T cells targeting CEA-positive carcinoma. Frontiers in Immunology, 14, 1182409.
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3

Generation and Characterization of 19BBz CAR T Cells

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19BBz CAR T cells were obtained from Unum Therapeutics. 19BBz T cells were generated using the commercial vector pLenti6.4/R4R2/V5-DEST (Invitrogen) with an EF1α promoter added and the PGK promoter and blasticidin S deaminase genes removed. The CD19 CAR was cloned downstream of the EF1α promoter. The 19BBz CAR was previously described53 and is composed of a CD8α signal peptide, a CD19-targeting scFv, a CD8α hinge and transmembrane domain, a 4-1BB costimulatory domain, and a CD3ζ signaling domain. The CD19 CAR protein sequence is as follows: MALPVTALLLPLALLLHAARPDIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEITGGGGSGGGGSGGGGSEVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSSTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR. 19BBz T cells were polyclonally re-stimulated with APC-ms or Dynabeads as described above. CAR expression was evaluated by primary staining with biotin-Protein L (Genscript) and secondary staining with APC-streptavidin (Biolegend).
Cheung A.S., Zhang D.K., Koshy S.T, & Mooney D.J. (2018). Scaffolds that mimic antigen-presenting cells enable ex vivo expansion of primary T-cells. Nature biotechnology, 36(2), 160-169.
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4

Generation and Characterization of 19BBz CAR T Cells

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19BBz CAR T cells were obtained from Unum Therapeutics. 19BBz T cells were generated using the commercial vector pLenti6.4/R4R2/V5-DEST (Invitrogen) with an EF1α promoter added and the PGK promoter and blasticidin S deaminase genes removed. The CD19 CAR was cloned downstream of the EF1α promoter. The 19BBz CAR was previously described53 and is composed of a CD8α signal peptide, a CD19-targeting scFv, a CD8α hinge and transmembrane domain, a 4-1BB costimulatory domain, and a CD3ζ signaling domain. The CD19 CAR protein sequence is as follows: MALPVTALLLPLALLLHAARPDIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEITGGGGSGGGGSGGGGSEVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSSTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR. 19BBz T cells were polyclonally re-stimulated with APC-ms or Dynabeads as described above. CAR expression was evaluated by primary staining with biotin-Protein L (Genscript) and secondary staining with APC-streptavidin (Biolegend).
Cheung A.S., Zhang D.K., Koshy S.T, & Mooney D.J. (2018). Scaffolds that mimic antigen-presenting cells enable ex vivo expansion of primary T-cells. Nature biotechnology, 36(2), 160-169.
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