Log In Sign up
The largest database of trusted experimental protocols

Beta mark tcr vβ repertoire kit

Manufactured by Beckman Coulter
Visit Supplier
Sourced in United States

The Beta Mark TCR Vβ Repertoire Kit is a laboratory product designed for the analysis of T-cell receptor (TCR) Vβ repertoire. It provides a tool for researchers to quantify and characterize the diversity of the T-cell Vβ chain within a sample.

Automatically generated - may contain errors

Lab products found in correlation

Navios, by Beckman Coulter (3 mentions) Immunoseq, by Adaptive Biotechnologies (1 mentions) Kaluza analysis software, by Beckman Coulter (1 mentions) Dneasy blood and tissue kit, by Qiagen (1 mentions) Anti cd56, by Beckman Coulter (1 mentions) Anti cd8 apc h7 sk1, by BD (1 mentions) Pe anti cd107a h4a3, by BD (1 mentions) Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions) Interleukin 2 (il 2), by Novartis (1 mentions) 7 aminoactinomycin d, by BD (1 mentions) Collagenase, by Merck Group (1 mentions) Cytostim, by Miltenyi Biotec (1 mentions) Cfda se, by Thermo Fisher Scientific (1 mentions) X vivo 15 media, by Lonza (1 mentions) Acd28, by Miltenyi Biotec (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Hyaluronidase, by Merck Group (1 mentions) Dnase, by Merck Group (1 mentions) Accuri c6 flow cytometer, by BD (1 mentions) Fixation buffer, by BioLegend (1 mentions)

14 protocols using beta mark tcr vβ repertoire kit

1

Profiling PBMC Immunophenotype and TCR

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell surface markers of peripheral blood mononuclear cells (PBMCs), lymphocyte proliferative response to mitogens, and the amount of T-cell receptor excision circles (TRECs) were determined as previously described (23 (link)). The analysis of T cell receptor (TCR) Vβ expression were determined according to manufacturer's manual (Beta Mark TCR Vβ Repertoire Kit, Beckman Coulter).
Lev A., Simon A.J., Barel O., Eyal E., Glick-Saar E., Nayshool O., Birk O., Stauber T., Hochberg A., Broides A., Almashanu S., Hendel A., Lee Y.N, & Somech R. (2019). Reduced Function and Diversity of T Cell Repertoire and Distinct Clinical Course in Patients With IL7RA Mutation. Frontiers in Immunology, 10, 1672.
+ Open protocol
+ Expand
2

Analyzing T-cell Repertoire in CAR T-cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Genomic DNA from pre-infusion CART-PSMA-TGFβRDN cells and sorted post-infusion CAR T-cells was isolated using the DNeasy Blood and Tissue Kit (Qiagen). Initial flow cytometric assessment the T-cell repertoire by flow cytometry was conducted using the Beta Mark TCRVβ repertoire kit and Kaluza analysis software (Beckman Coulter Life Sciences). Subsequent TCRVβ deep sequencing was carried out by immunoSEQ (Adaptive Biotechnologies). Only productive TCR rearrangements were used in the evaluation of clonotype frequencies.
Narayan V., Barber-Rotenberg J.S., Jung I.Y., Lacey S.F., Rech A.J., Davis M.M., Hwang W.T., Lal P., Carpenter E.L., Maude S.L., Plesa G., Vapiwala N., Chew A., Moniak M., Sebro R.A., Farwell M.D., Marshall A., Gilmore J., Lledo L., Dengel K., Church S.E., Hether T.D., Xu J., Gohil M., Buckingham T.H., Yee S.S., Gonzalez V.E., Kulikovskaya I., Chen F., Tian L., Tien K., Gladney W., Nobles C.L., Raymond H., Hexner E.O., Siegel D.L., Bushman F.D., June C.H., Fraietta J.A, & Haas N.B. (2022). PSMA-targeting TGFβ-insensitive Armored CAR T-cells in Metastatic Castration Resistant Prostate Cancer: a Phase 1 Trial. Nature medicine, 28(4), 724-734.
+ Open protocol
+ Expand
3

Lymphocyte Subset Profiling by Flow Cytometry

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell surface markers of peripheral blood mononuclear cells were measured by flow cytometry (NAVIOS; Beckman Coulter) with immunofluorescent staining using anti-CD3, anti-CD4, anti CD8, anti-CD19, anti-CD16, and anti-CD56 antibodies (Beckman Coulter). T-cell receptor (TCR) v-β expression was determined as directed by the manufacturer (Beta Mark TCR v-β Repertoire Kit; Beckman Coulter). Reference range for lymphocyte subsets in adults were taken from Apoil et al. (15 (link))
Shamriz O., Zahalka N., Simon A.J., Lev A., Barel O., Mor N., Tal Y., Segel M.J., Somech R., Yonath H, & Toker O. (2022). GATA2 Deficiency in Adult Life Is Characterized by Phenotypic Diversity and Delayed Diagnosis. Frontiers in Immunology, 13, 886117.
+ Open protocol
+ Expand
4

Lymphocyte Proliferation and NK Cell Degranulation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Lymphocyte proliferative response to mitogens and the amount of TCR excision circles were determined as previously described (Amariglio et al., 2010 (link)). Proliferative response was measured by labeling PBMCs with 2.5 mM CFSE (Thermo Fisher Scientific), 7-aminoactinomycin D (BD Biosciences), PacB anti-CD3 (SK7; Biolegend), PE anti-CD25 (M-A251; BD Biosciences), APC-H7 anti-CD8 (SK1; BD Biosciences), and BV711 anti-CD4 (SK3; BD Biosciences) 4 d after stimulation. Activation response was determined by CD25 levels in stimulated PBMCs. The analysis of TCR Vβ expression was determined according to manufacturer’s manual (Beta Mark TCR Vβ Repertoire Kit; Beckman Coulter). NK degranulation was assessed by CD107a surface staining without stimulation (medium-cultured cells) and 3 h after stimulation with K562 cells at a ratio of 1:1 as previously described (Bryceson et al., 2012 (link)). The erythroleukemic cell line K562 (CCL-243; ATCC) was used as a target cell line. NK cells were cultured in medium containing 600 U/ml IL-2 (Novartis) for 48 h to assess the degranulation of activated NK cells. For surface staining, the following antibodies were used: PacB anti-CD3 (SK7; Biolegend), APC-H7 anti-CD8 (SK1; BD Biosciences), FITC anti-CD56 (NCAM16.2; BD Biosciences), and PE anti-CD107a (H4A3; BD Biosciences).
Lev A., Lee Y.N., Sun G., Hallumi E., Simon A.J., Zrihen K.S., Levy S., Beit Halevi T., Papazian M., Shwartz N., Somekh I., Levy-Mendelovich S., Wolach B., Gavrieli R., Vernitsky H., Barel O., Javasky E., Stauber T., Ma C.A., Zhang Y., Amariglio N., Rechavi G., Hendel A., Yablonski D., Milner J.D, & Somech R. (2020). Inherited SLP76 deficiency in humans causes severe combined immunodeficiency, neutrophil and platelet defects. The Journal of Experimental Medicine, 218(3), e20201062.
+ Open protocol
+ Expand
5

Comprehensive Immunological Assay Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
RPMI 1640 was from Lifetechnologies. X-Vivo15 media was supplied by Lonza. aCD3, aCD4, aCD8 (recognizing the α chain), aCD28, aCD45RO, aCD56, aCCR7, a-IL17, aTCRα24-Jα18 (clone: 6B11), cytokine secretion assays for IFNγ and IL-4, a-fibroblast microbeads and Cytostim were purchased from Miltenyi. Collagenase, Hyaluronidase and DNAse were all from Sigma-Aldrich. aCD45 and aCD38 were from Immunotools. CFDA-SE was purchased from Molecular Probes/Invitrogen. Intra staining Kit, aCD16, aCD8β and aCD3 were from Beckton Dickinson. aCXCR5 was supplied by R&D Systems and aIL21 was from ebioscience. The Beta Mark TCRVβ Repertoire Kit was supplied by Beckman Coulter. The antibodies were used in different conjugates of FITC, PE, PerCp, APC, APC-Vio770 and PE-Cy7.
Quandt D., Rothe K., Scholz R., Baerwald C.W, & Wagner U. (2014). Peripheral CD4CD8 Double Positive T Cells with a Distinct Helper Cytokine Profile Are Increased in Rheumatoid Arthritis. PLoS ONE, 9(3), e93293.
+ Open protocol
+ Expand
6

TCR-Vβ Profiling in Patient PBMCs

Check if the same lab product or an alternative is used in the 5 most similar protocols
The surface expression of T-cell receptors (TCRs) with variable gene from the β-chain in PBMCs isolated from patients’ blood were determined and quantified using flow cytometry (NAVIOS, Beckman Coulter). The analysis of TCR-Vβ expression were determined according to manufacturer’s manual (Beta Mark TCR-Vβ repertoire kit, Beckman Coulter). We compared the results to healthy control values provided by the kit (n = 58).
Palevski D., Simon A., Lev A., Somech R, & Lee Y.N. (2022). Inadequate Activation of γδT- and B-cells in Patient with Wiskott-Aldrich Syndrome (WAS) Portrayed by TRG and IGH Repertoire Analyses. Journal of Clinical Immunology, 43(1), 109-122.
+ Open protocol
+ Expand
7

Profiling T Cell Receptor Diversity

Check if the same lab product or an alternative is used in the 5 most similar protocols
The TCR Vβ repertoire diversity of the expanded T cells was determined using a panel of 24 TCR Vβ-specific antibodies (Beta Mark TCR Vβ Repertoire Kit; IM3497; Beckman Coulter). After staining, cells were analyzed with a C6 Accuri Flow Cytometer (BD Biosciences).
Nicolas P., Ollier J., Mori D., Voisinne G., Celis-Gutierrez J., Gregoire C., Perroteau J., Vivien R., Camus M., Burlet-Schiltz O., Gonzalez de Peredo A., Clémenceau B., Roncagalli R., Vié H, & Malissen B. (2022). Systems-level conservation of the proximal TCR signaling network of mice and humans. The Journal of Experimental Medicine, 219(2), e20211295.
+ Open protocol
+ Expand
8

Comprehensive Cell Surface and Intracellular Staining

Check if the same lab product or an alternative is used in the 5 most similar protocols
For surface staining, cells were washed in PBS supplemented with 2% fetal bovine serum and stained with the appropriate antibodies for 30 min at 4 °C in the dark. The dilutions of antibodies are provided in Supplementary Table 3. After the surface staining, the cells were washed, and 1 µg/ml propidium iodide (Sigma-Aldrich) was added to exclude dead cells. To access the TCR Vβ repertoire, the Beta Mark TCR Vβ Repertoire Kit (IM3497, Beckman Coulter) was used. For intracellular staining, reagents (Fixation Buffer, Cell Staining Buffer, and Permeabilization Buffer) and True-Nuclear Transcription Factor Buffer set purchased from BioLegend were used. Data were acquired on an LSRFortessa (BD) or FACSAria flow cytometer (BD) and analyzed with FlowJo software (Tree Star).
Ito T., Kawai Y., Yasui Y., Iriguchi S., Minagawa A., Ishii T., Miyoshi H., Taketo M.M., Kawada K., Obama K., Sakai Y, & Kaneko S. (2021). The therapeutic potential of multiclonal tumoricidal T cells derived from tumor infiltrating lymphocyte-derived iPS cells. Communications Biology, 4, 694.
+ Open protocol
+ Expand
9

Multiparameter Flow Cytometry Immunophenotyping

Check if the same lab product or an alternative is used in the 5 most similar protocols
High sensitivity multiparameter flow cytometric immunophenotyping was performed with a FACS Canto 10-color instrument (BD Biosciences, San Jose, CA) using the following antibodies: CD45-V500C (2D1, BD Biosciences), CD3-PC7 (UCHT1, Beckman-Coulter, Miami, FL), CD2-BV421 (RPA-2.10, BD Horizon, San Jose, CA), CD5-APC (L17F12, BD Biosciences), CD7-BB515 (M-T701, BD Horizon), CD4-APC-A700 (13B8.2, Beckman-Coulter), CD8-APC-H7 (SK1, BD Biosciences), CD56-PE (N901, Beckman-Coulter), CD26-PerCP-Cy5.5 (BA5B, BioLegend, San Diego, CA) and CD279 (PD-1)-BV605 (EH12.2H7, BioLegend). At least 200,000 cells were acquired and at least 20 events were required for a positive result. The data was analyzed using custom software (“Woodlist,” gift of B.L. Wood, University of Washington, Seattle, WA). Abnormal T cell populations were identified by visual assessment based on aberrant antigen expression. Median fluorescent intensity (MFI) of markers was evaluated. Abnormal populations were considered “dim” or “bright” for a given marker if the fluorescent intensity was ≥1/3 log different from that of corresponding internal normal CD4+ T cells. In select cases, clonality of T cell populations was assessed with a Beta Mark TCR Vβ Repertoire Kit (Beckman-Coulter) using a modified, single-tube method as described previously (25 (link)).
Lewis N.E., Gao Q., Petrova-Drus K., Pulitzer M., Sigler A., Baik J., Moskowitz A.J., Horwitz S.M., Dogan A, & Roshal M. (2022). PD-1 improves accurate detection of Sezary cells by flow cytometry in peripheral blood in mycosis fungoides/Sezary syndrome. Cytometry. Part B, Clinical cytometry, 102(3), 189-198.
+ Open protocol
+ Expand
10

Degranulation and Apoptosis Assays for Engineered T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Antibodies used in this study are listed in online supplemental table S1. Cells were stained according to the manufacturer’s instructions. To detect the expression of the lysosomal-associated membrane protein 1 (LAMP1/CD107a) (as a surrogate marker for degranulation) on the surface of CD8+ T-cells, effector cells were incubated for 24 hours with target tumor cells in the presence of monensin (Monensin Solution 1000X, eBioscience) and the CD107a antibody. Phorbol 12-myristate 13-acetate (PMA) + ionomycin (Sigma-Aldrich) were used as control inducers of CD107a expression. The determination of apoptotic cells was performed with a standard Annexin V and propidium iodide (PI) (BD Biosciences) staining according to the manufacturer’s instructions. TCR Vβ repertoire of TCR-engineered T cells in vivo was analyzed by flow cytometry with antibodies from the Beta Mark TCR Vβ Repertoire Kit (Beckman Coulter) directed against 19 individual TCR/Vβ chains. Flow cytometry acquisitions were performed on a FACSCanto II and BD FACSLyric (BD Biosciences), and data were analyzed with FlowJo_V10 software (Tree Star).
Legscha K.J., Antunes Ferreira E., Chamoun A., Lang A., Awwad M.H., Ton G.N., Galetzka D., Guezguez B., Hundemer M., Bourdon J.C., Munder M., Theobald M, & Echchannaoui H. (2021). Δ133p53α enhances metabolic and cellular fitness of TCR-engineered T cells and promotes superior antitumor immunity. Journal for Immunotherapy of Cancer, 9(6), e001846.
+ 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?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!