Trucount absolute count tubes

Manufactured by BD

Sourced in United States

The BD Trucount Absolute Count Tubes are laboratory equipment used to determine the absolute count of specific cell populations in a sample. They provide a standardized method for quantifying the number of cells per unit volume, enabling accurate and reproducible cell counting.

Automatically generated - may contain errors

Lab products found in correlation

Opteia human elisa kit, by BD (2 mentions) Lsrfortessa, by BD (2 mentions) Facs lysing solution, by BD (2 mentions) Accuri c6 plus, by BD (2 mentions) Facscanto 2, by BD (1 mentions) Cd45 fitc, by BD (1 mentions) Versant hiv 1 rna 3.0 assay, by Siemens (1 mentions) Ficoll hypaque density gradient centrifugation, by Cytiva (1 mentions) Facscalibur flow cytometer, by BD (1 mentions) Facscanto 2 flow cytometry system, by BD (1 mentions) Cd56 pe, by BD (1 mentions) Cd19 apc, by BD (1 mentions) Facsdiva software, by BD (1 mentions) Cd3 fitc, by BD (1 mentions) Cd4 pe cy7, by BD (1 mentions) Cd8 apc cy7, by BD (1 mentions) Cd45 percp cy5, by BD (1 mentions) Onecomp ebeads, by Thermo Fisher Scientific (1 mentions) Anti cd3, by Thermo Fisher Scientific (1 mentions) Facscanto 2 flow cytometer, by BD (1 mentions)

9 protocols using trucount absolute count tubes

Splenic Leukocyte Immunophenotyping

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

Fresh spleens were minced and dissolved in PBS, filtered through a 40-μm cell strainer, and centrifuged at 350 ×g for 10 min at 4°C. Cells were washed with PBS and re-suspended in 2 ml PBA (∼1–5 × 10⁷ cells/ml). The suspension (50 μl) was incubated with anti-CD3 (eBioscience, San Diego, CA, United States PE-Cyanine7, 1:100), CD4 (eBioscience, San Diego, CA, United States APC/Cy7, 1:100), CD8 (eBioscience, San Diego, CA, United States APC, 1:100), CD19 (eBioscience, San Diego, CA, United States PE, 1:100) for 20 min at 4°C, and then mixed with 448 μl PBS. The suspension was transferred to BD Trucount Absolute Count Tubes (BD, New Jersey, FL). The standard of the experiment is to collect 4000 beads in every BD Trucount Absolute Count Tubes to stop counting. Cells and beads were analyzed by using a BD FacScanto II flow cytometer.
Non-specific binding of secondary antibodies was quantified, and a fluorescent signal was subtracted from values of experimental groups. Single-stained cells or OneComp eBeads (eBioscience, San Diego, CA, United States) were used for calculations. Unstained cells, GFP-expressing cells with 7-AAD, and fluorescence minus one (FMO) controls were used for cytometry and gating set up.

Hu Z., Cui Y., Qiao X., He X., Li F., Luo C., Wang S., Li C, & Dai R. (2018). Silencing miR-150 Ameliorates Experimental Autoimmune Encephalomyelitis. Frontiers in Neuroscience, 12, 465.

+ Open protocol

+ Expand

Isolation and Purification of CD34+ Cells from Cord Blood

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

Fresh cord blood (CB) samples were collected after normal vaginal delivery in women without infectious diseases or maternal complications, after informed written consent. Mononuclear cells (MNC) were isolated from 30 ml of CB after density gradient centrifugation using lymphocytes separation media (Histopaque^®, Sigma-Aldrich) stratification (2000 rpm, 20 min). After washing with Phosphate-Buffered Saline (PBS) (Gibco), MNC were incubated with immunomagnetic beads (CD34+) (Miltenyi) following manufacturer’s instruction. After CD34+ cell recovery, cell purity was assessed by flow cytometry (FACS Canto II^®, BD) using a panel of CD34-PE and CD45-FITC (BD) fluorochrome-conjugated monoclonal antibodies. TruCount^® Absolute-Count Tubes (BD) were used for determining the number of CD34+ cells. The strategy of gating is reported in the S1 and S2 Figs. CB samples were processed within 8 hours from collection; cells with at least 90% of purity were used for co-culture experiments.

Perucca S., Di Palma A., Piccaluga P.P., Gemelli C., Zoratti E., Bassi G., Giacopuzzi E., Lojacono A., Borsani G., Tagliafico E., Scupoli M.T., Bernardi S., Zanaglio C., Cattina F., Cancelli V., Malagola M., Krampera M., Marini M., Almici C., Ferrari S, & Russo D. (2017). Mesenchymal stromal cells (MSCs) induce ex vivo proliferation and erythroid commitment of cord blood haematopoietic stem cells (CB-CD34+ cells). PLoS ONE, 12(2), e0172430.

+ Open protocol

+ Expand

PBMC Isolation and Viral Load Measurement

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

Forty ml of whole blood were collected from study participants, centrifuged to separate plasma, and stored at −80°C. Peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll-Hypaque density gradient centrifugation (Amersham, Sweden) and cryopreserved for subsequent functional assays. Plasma viral load (VL) was determined by branched-DNA, Versant HIV-1 RNA 3.0 assay (Siemens Healthcare, UK). CD4⁺ and CD8⁺ T-cell counts were determined using TruCount absolute-count tubes (BD Bioscences, USA) on a BD FACSCalibur flow cytometer.

Salido J., Ruiz M.J., Trifone C., Figueroa M.I., Caruso M.P., Gherardi M.M., Sued O., Salomón H., Laufer N., Ghiglione Y, & Turk G. (2018). Phenotype, Polyfunctionality, and Antiviral Activity of in vitro Stimulated CD8+ T-Cells From HIV+ Subjects Who Initiated cART at Different Time-Points After Acute Infection. Frontiers in Immunology, 9, 2443.

+ Open protocol

+ Expand

Cytotoxicity Assay of CAR T-cells against B-ALL

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

For cytotoxicity assays, BM-MSCs were seeded at 1×10⁴ cells/well in flat-bottom 96-well plates containing complete Advanced DMEM medium and incubated at 37°C for 24 hours. The next day, target cells (B-ALL cell lines or primary B-ALL cells, 1×10⁵ target cells/well) were incubated with CD19-CAR T-cells at an Effector:Target (E:T) ratio of 1:1, or alone, for the indicated periods in the absence/presence of BM-MSC. Cell lines were cultured in RPMI complete medium and primary cells were cultured in StemSpan supplemented with 20% heat-inactivated FBS, P/S, ITS, hSCF (100 ng/mL), hFLT3-L (100 ng/mL), hIL-3 (10 ng/mL) and hIL-7 (10 ng/mL). After 2 and 6 days of culture, non-adherent cells were collected, washed and stained with anti-CD3, anti-CD19, anti-CD10, anti-CD22 mAbs and 7-AAD. CAR T-cell-mediated cytotoxicity was determined by analyzing the residual living (CD10⁺CD22⁺7-AAD^-) target cells at each time point. BD TruCount absolute count tubes (BD Biosciences) were used for absolute cell counting. Quantification of the pro-inflammatory cytokines IL-2, IFN-γ and TNF-α was measured by ELISA using the OptEIA Human ELISA Kit (BD Biosciences) on supernatants harvested after 2 days of coculture at a 1:1 E:T ratio. ELISA determinations were performed in triplicate.

Zanetti S.R., Romecin P.A., Vinyoles M., Juan M., Fuster J.L., Cámos M., Querol S., Delgado M, & Menendez P. (2020). Bone marrow MSC from pediatric patients with B-ALL highly immunosuppress T-cell responses but do not compromise CD19-CAR T-cell activity. Journal for Immunotherapy of Cancer, 8(2), e001419.

+ Open protocol

+ Expand

Comprehensive Immune Cell Profiling

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

The proportions of neutrophils, lymphocytes, monocytes, CD4⁺ T, CD8⁺ T, CD19⁺ B, NK (CD3^-CD56⁺), and NKT (CD3⁺CD56⁺) cell were analyzed by flow cytometry. The absolute counts of immune cells were measured by using BD TruCount Absolute-Count Tubes (BD340334). The following antibodies (CD3-FITC, CD8-APC-Cy7, CD4-PE-Cy7, CD45-Percp-Cy5.5, CD56-PE, and CD19-APC) were used, and all reagents were purchased from BD Biosciences. All samples were detected by a BD FACS Canto II flow cytometry system and analyzed with the BD FACS Diva software.

Liao B., Liu Z., Tang L., Li L., Gan Q., Shi H., Jiao Q., Guan Y., Xie M., He X., Zhao H., Chen W., Liu Y., Li L., Wang Y., Cao Y., Shi Y., Li Y, & Lei C. (2021). Longitudinal clinical and radiographic evaluation reveals interleukin-6 as an indicator of persistent pulmonary injury in COVID-19. International Journal of Medical Sciences, 18(1), 29-41.

+ Open protocol

+ Expand

Evaluating CAR T-cell Cytotoxicity

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

Target cells (B-ALL cell lines, primary B-ALL cells, and B-ALL PDXs cells; 1 × 10⁵ target cells/well) were incubated with Mock-, CD19-, or Tan-CAR T cells at different E:T ratios for the indicated time periods. Cell lines were cultured in complete RPMI medium and primary cells/PDXs were cultured in StemSpan SFEM supplemented with 20% heat-inactivated FBS, P/S, ITS, hSCF (100 ng/mL), hFLT3-L (100 ng/mL), hIL-3 (10 ng/mL), and hIL-7 (10 ng/mL). At each time point, cells were collected, washed, and stained with anti-CD3, anti-CD19, anti-CD10, and anti-CD22 mAbs, and 7-AAD. CAR T cell-mediated cytotoxicity was determined by analyzing the residual living (7-AAD⁻CD3⁻GFP⁻CD10⁺) target cells at each time point and E:T ratio (Figure S1A). BD TruCount absolute count tubes (BD Biosciences) were used for absolute cell counting. The quantification of IL-2, IFN-γ, and TNF-α was measured by enzyme-linked immunosorbent assay (ELISA) using the OptEIA Human ELISA Kit (BD Biosciences) on supernatants harvested after 1–2 days of co-culture at a 1:1 E:T ratio. ELISA determinations were performed in triplicate.

Zanetti S.R., Velasco-Hernandez T., Gutierrez-Agüera F., Díaz V.M., Romecín P.A., Roca-Ho H., Sánchez-Martínez D., Tirado N., Baroni M.L., Petazzi P., Torres-Ruiz R., Molina O., Bataller A., Fuster J.L., Ballerini P., Juan M., Jeremias I., Bueno C, & Menéndez P. (2021). A novel and efficient tandem CD19- and CD22-directed CAR for B cell ALL. Molecular Therapy, 30(2), 550-563.

+ Open protocol

+ Expand

Rhesus Macaque T-Cell Enumeration

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

Blood samples from rhesus macaques were all collected following standard protocols. BD Trucount Absolute Count Tubes (BD, Cat. No. 340334) were used to quantify the T cells in 100 µL of blood. Antibodies that label different kinds of T cells were adopted in this study. Information on antibodies is listed in Table S3. FACS Lysing solution (BD Cat. No. 349202) was diluted 10 times before being added, and the suspensions were incubated at 4°C for 10 min. All data were collected through BD Accuri C6 Plus. PBMCs were collected to test the phenotypic characteristics of T cells, and the antibodies used are all listed in Table S3. All data were collected by an LSRFortessa (BD Biosciences) instrument and were analyzed using FlowJo software (Tree Star, Inc).

He Y., Wu C., Liu Z., Zhang Y., Feng F., Lin Z., Wang C., Yang Q., Wen Z., Liu Y., Zhang F., Lin Y., Zhang H., Qu L., Li L., Cai W., Sun C., Chen L, & Li P. (2023). Arsenic trioxide-induced apoptosis contributes to suppression of viral reservoir in SIV-infected rhesus macaques. Microbiology Spectrum, 11(5), e00525-23.

+ Open protocol

+ Expand

Rhesus Macaque T-Cell Enumeration

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

+ Open protocol

+ Expand

Quantification of T Cell Subsets

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

The numbers of circulating CD4⁺ T and CD8⁺ T lymphocytes were determined using BD Trucount absolute count tubes according to the manufacturer’s instructions (BD Biosciences). For detection of central memory CD4⁺ T (CD3 ⁺CD4 ⁺CD28 ⁺CD95 ⁺cells), effector memory CD4⁺ T (CD3 ⁺CD4 ⁺CD28^-CD95⁺ cells) and TFH cell (CD3 ⁺CD4 ⁺CXCR5 ⁺PD-1⁺) cells, PBMCs were stained with the following fluorescein-labeled antibodies: anti-CD3 (BD Bioscience), anti-CD4 (BD Bioscience), anti-CD8 (BD Bioscience), anti-CD28 (BD Bioscience), anti-CD95 (BD Bioscience), anti-CXCR5 (eBioscience), and anti-PD-1 (eBioscience) for 30 min and detected with a BD FACS LSR Fortessa flow cytometer (BD Biosciences, USA). Data were analyzed using FlowJo software (Tree Star, USA).

Li P., Wang Q., He Y., Yang C., Zhang Z., Liu Z., Liu B., Yin L., Cui Y., Hu P., Liu Y., Zheng P., Wang W., Qu L., Sun C., Guan S., Feng L, & Chen L. (2023). Booster vaccination is required to elicit and maintain COVID-19 vaccine-induced immunity in SIV-infected macaques. Emerging Microbes & Infections, 12(1), e2136538.

+ 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!