Log In Sign up
The largest database of trusted experimental protocols

Cd3 af700 clone ucht1

Manufactured by BD
Visit Supplier

The CD3-AF700 (clone UCHT1) is a fluorochrome-conjugated monoclonal antibody that binds to the CD3 antigen, a component of the T cell receptor complex. It is commonly used in flow cytometry applications to identify and quantify T cells in biological samples.

Automatically generated - may contain errors

Lab products found in correlation

Lsr 2 flow cytometer, by BD (4 mentions) Cd4 apc h7 clone rpa t4, by BD (3 mentions) Alexa fluor dye coupled neutravidin beads, by Thermo Fisher Scientific (2 mentions) Cd45 buv395 clone hi30, by BD (2 mentions) Cd8 apc h7 clone sk1, by BD (2 mentions) Pd 1 bv421 clone eh12.2h7, by BioLegend (2 mentions) Fortessa flow cytometer, by BD (2 mentions) Recombinant ebov gp δtm, by IBT Bioservices (2 mentions) Fix and perm, by Thermo Fisher Scientific (2 mentions) Golgistop, by Thermo Fisher Scientific (2 mentions) Maxisorp 96 well plate, by Thermo Fisher Scientific (2 mentions) Lsr fortessa cytometer, by BD (1 mentions) Transcription factor buffer set, by BD (1 mentions) Cd8 percp cy5.5 clone sk1, by BioLegend (1 mentions) Absolute count microsphere, by Thermo Fisher Scientific (1 mentions) Tigit pedazzle594, by BioLegend (1 mentions) Lysis buffer, by BD (1 mentions) Fixable viability stain 620, by BD (1 mentions) Anti cd8a allophycocyanin fire 750 clone rpa t8, by BioLegend (1 mentions) Cd69 apc, by BioLegend (1 mentions)

9 protocols using cd3 af700 clone ucht1

1

Zika Virus Antibody Phagocytosis Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Recombinant soluble ZIKV E protein was biotinylated and coupled to Alexa Fluor dye coupled Neutravidin beads (Life Technologies). Microscale-purified mAbs were tested at a single 1:10 dilution; concentrations were not normalized. Antibodies were diluted in cell culture medium and incubated with beads for 2 hrs at 37°C. White blood cells were isolated from the peripheral blood of subjects by lysis of red blood cells, followed by three washes with PBS. Cells were added at a concentration of 5.0 × 104 cells/well and incubated for 1 hr at 37°C. Cells were stained with anti-human CD66b (Pacific Blue, Clone G10F5; BioLegend), -CD3 (AF700, Clone UCHT1; BD Biosciences), and -CD14 (APC-Cy7, Clone MφP9; BD Biosciences) antibodies (1:100 dilution of each), then fixed with 4% PFA, and analyzed by flow cytometry on a BD LSR II flow cytometer. Neutrophils were defined as SSC-Ahigh CD66b+, CD3-, CD14-. A phagocytic score was determined using the percentage of Alexa Fluor 488+ cells and the MFI of the Alexa Fluor 488+ cells. Z-score values were calculated as described above. The previously identified mAb ZIKV-117 was used as a positive control, and mAb FLU-5J8 was used as a negative control.
Gilchuk P., Bombardi R.G., Erasmus J.H., Tan Q., Nargi R., Soto C., Abbink P., Suscovich T.J., Durnell L.A., Khandhar A., Archer J., Liang J., Fouch M.E., Davidson E., Doranz B.J., Jones T., Larson E., Ertel S., Granger B., Fuerte-Stone J., Roy V., Broge T., Linnekin T.C., Linde C.H., Gorman M.J., Nkolola J., Alter G., Reed S.G., Barouch D.H., Diamond M.S., Crowe JE J.r., Van Hoeven N., Thackray L, & Carnahan R. (2020). Integrated pipeline for the accelerated discovery of antiviral antibody therapeutics. Nature biomedical engineering, 4(11), 1030-1043.
+ Open protocol
+ Expand
2

Flow Cytometry Analysis of B-ALL and Healthy Samples

Check if the same lab product or an alternative is used in the 5 most similar protocols
Flow cytometry was performed for three newly diagnosed and untreated B-ALL patients and three healthy individuals. The clinical information of these samples is listed in Supplementary Table S7. Cell surface staining and intracellular staining were carried out in 1× phosphate buffered saline (PBS) and using the BD Transcription Factor Buffer Set, respectively. Flow cytometry was performed using the BD LSRFortessa cytometer, and data analysis was performed using FlowJo (version 10.5.3) (BD). Cell surface and intracellular staining was performed using the following fluorophore-conjugated antibodies: CD45-BUV395 (clone HI30, BD), CD3-AF700 (clone UCHT1, BD), CD4-APC-H7 (clone RPAT4, BD), CD8-Percp-cy5.5 (clone SK1, BioLegend), Ki-67-PE-CF594 (clone Ki-67, BioLegend), PD-1-BV421 (clone MIH4, BD), and CD103-BB515 (clone Ber-ACT8, BD). Cell surface and intracellular staining was performed according to the manufacturer’s instructions.
Lai W., Wang X., Liu L., Xu L., Mao L., Tan J., Zha X., Zhan H., Lei W., Lan Y., Chen G., Li Y, & Luo O.J. (2022). Single-cell profiling of T cells uncovers a tissue-resident memory-like T-cell subset associated with bidirectional prognosis for B-cell acute lymphoblastic leukemia. Frontiers in Immunology, 13, 957436.
+ Open protocol
+ Expand
3

Multiparameter Flow Cytometry for Cell Phenotyping

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell surface staining for flow cytometry was performed using the following antibodies: CD45-BUV395 (clone HI30, BD), CD3-AF700 (clone UCHT1, BD), CD4-APC-H7 (clone RPA-T4, BD), CD8-APC-H7 (clone SK1, BD), TIGIT-PE/Dazzle™ 594 (clone A15153G, Biolegend), TIGIT-BV421 (clone A15153G, Biolegend), and PD1-BV421 (clone EH12.2H7, Biolegend). These antibodies were used to analyze surface receptors in two different panels. Isotype-matched antibodies, labeled with the proper fluorochromes, were used as negative controls. Extracellular staining was performed according to the manufacturer’s instructions. Briefly, an antibody cocktail was added to whole blood or BM samples, which were then incubated at room temperature for 15 minutes in the dark, followed by red cell lysis with lysis buffer (BD; Cat: 555899). The lysed cells were washed twice with phosphate buffer saline (PBS), and then 350 μl stain buffer was added for further flow cytometer analysis. Twenty microliters of absolute count microsphere (Thermo; Cat: C36950) were added to the samples for absolute number analysis. Cells were analyzed using a BD Fortessa flow cytometer (BD Biosciences), and data analysis performed with Flowjo 10.6 software.
Xu L., Liu L., Yao D., Zeng X., Zhang Y., Lai J., Zhong J., Zha X., Zheng R., Lu Y., Li M., Jin Z., Hebbar Subramanyam S., Chen S., Huang X, & Li Y. (2021). PD-1 and TIGIT Are Highly Co-Expressed on CD8+ T Cells in AML Patient Bone Marrow. Frontiers in Oncology, 11, 686156.
+ Open protocol
+ Expand
4

Flow cytometry cell surface staining

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell surface staining for flow cytometry was performed using the following antibodies: CD3-AF700 (clone UCHT1, BD), CD4-APC-H7 (clone RPA-T4, BD), CD8-APC-H7 (clone SK1, BD) and PD-1-BV421 (clone EH12.2H7, Biolegend). Isotype-matched antibodies, labeled with the proper fluorochromes, were used as negative controls. Cells were analyzed using a BD Fortessa flow cytometer (BD Biosciences), and data analysis was performed with Flowjo 10.6 software as previously described (22 (link)).
Geng S., Xu R., Huang X., Li M., Deng C., Lai P., Wang Y., Wu P., Chen X., Weng J, & Du X. (2022). Dynamics of PD-1 expression are associated with treatment efficacy and prognosis in patients with intermediate/high-risk myelodysplastic syndromes under hypomethylating treatment. Frontiers in Immunology, 13, 950134.
+ Open protocol
+ Expand
5

Antigen-specific CD8+ T cell sorting

Check if the same lab product or an alternative is used in the 5 most similar protocols
Approximately 5 × 106 CD8+ T cells and 10 × 106 PBMCs were rested overnight and washed twice in PBS. The cells were then supplemented with 0.05 U/µL of RNase inhibitor (RNase outTM recombinant ribonuclease inhibitor) and stained with either PE-conjugated HLA B*57:01/ VSFIEFVGWL or HLA B*57:01/QSRGDENRGW tetramer at a final concentration of 10 nM for 20 min at room temperature in the dark. The cells were pelleted by centrifugation at 400 g for 7 min and washed in FACS buffer (PBS + 1%FBS), followed by surface staining with antibodies, fixable viability stain 620 (BD Biosciences, 1:1000 dilution), CD3-AF700 (clone UCHT1, BD Biosciences, 1:50 dilution), and anti-CD8a Allophycocyanin-Fire 750 (clone RPA-T8, BioLegend, 1:40 dilution) for 30 min at room temperature in the dark. After washing the cells twice in FACS buffer, cells were resuspended in 500 µL of FACS buffer and strained using a polystyrene tube with cell strainer cap (In Vitro Technologies) and supplemented with 0.1 U/µL RNase inhibitor. Cells were left on wet ice and protected from light until the time of sort. Finally, individual cells were sorted directly into each well of a 96-well plate containing 3 µL of lysis buffer using the FACSAria III cell sorter.
Sooda A., Rwandamuriye F., Wanjalla C.N., Jing L., Koelle D.M., Peters B., Leary S., Chopra A., Calderwood M.A., Mallal S.A., Pavlos R., Watson M., Phillips E.J, & Redwood A.J. (2022). Abacavir inhibits but does not cause self-reactivity to HLA-B*57:01-restricted EBV specific T cell receptors. Communications Biology, 5, 133.
+ Open protocol
+ Expand
6

Multiparameter Flow Cytometry Panel

Check if the same lab product or an alternative is used in the 5 most similar protocols
Fixable Viability Stain 510 was obtained from ThermoFisher Scientific (#L34957, 1/1000). The following antibodies were used in staining experiments: CD3 AF700 Clone UCHT1 (BD Biosciences, #557943, 1/50), CD4 PE-Cy7 Clone L200 (BD Bioscience, #560644, 1/200) and CD4 BV786 Clone SK3 (BD Biosciences, #563881, 1/200), CD8 BV510 Clone RPA-T8 (BioLegend, #301047, 1/200), CD45RO BV421 Clone UCHL1 (BD Biosciences, #562649, 1/100) or CD45RO PE Clone HI100 (BD Biosciences, #5555493, 1/20), CD27 BV605 Clone L128 (BD Biosciences, #562656, 1/100), PD1 BB700 Clone EH12.1 (BD Biosciences, #566461, 1/100), CD69 PerCP-Cy5.5 Clone FN50 (BD Biosciences, #560738) and CD69 APC (BioLegend, #310909, 1/50), CD25 BV421 Clone M-A251 (BD Biosciences, #562443, 1/50), HLA-DR FITC Clone REA805 (Miltenyi Biotech, #130-111-788, 1/50), GZMA PE-Cy7 Clone CB9 (BioLegend, #507221, 1/25), GZMB PB Clone GB11 (BioLegend, #515407, 1/50), CCL5 PerCP-Cy5.5 Clone VL1 (BioLegend, #515507, 1/50), CD127 PE-CF594 Clone HIL-7R-M21 (BD Biosciences, #562397, 1/50). For p24 staining, we used a combination of two antibodies: p24 KC57-FITC (Beckman Coulter, #6604665, 1/500) and p24 28B7-APC (MediMabs, #MM-0289-APC, 1/400).
Pardons M., Cole B., Lambrechts L., van Snippenberg W., Rutsaert S., Noppe Y., De Langhe N., Dhondt A., Vega J., Eyassu F., Nijs E., Van Gulck E., Boden D, & Vandekerckhove L. (2023). Potent latency reversal by Tat RNA-containing nanoparticle enables multi-omic analysis of the HIV-1 reservoir. Nature Communications, 14, 8397.
+ Open protocol
+ Expand
7

Zika Virus Antibody Phagocytosis Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Recombinant soluble ZIKV E protein was biotinylated and coupled to Alexa Fluor dye coupled Neutravidin beads (Life Technologies). Microscale-purified mAbs were tested at a single 1:10 dilution; concentrations were not normalized. Antibodies were diluted in cell culture medium and incubated with beads for 2 hrs at 37°C. White blood cells were isolated from the peripheral blood of subjects by lysis of red blood cells, followed by three washes with PBS. Cells were added at a concentration of 5.0 × 104 cells/well and incubated for 1 hr at 37°C. Cells were stained with anti-human CD66b (Pacific Blue, Clone G10F5; BioLegend), -CD3 (AF700, Clone UCHT1; BD Biosciences), and -CD14 (APC-Cy7, Clone MφP9; BD Biosciences) antibodies (1:100 dilution of each), then fixed with 4% PFA, and analyzed by flow cytometry on a BD LSR II flow cytometer. Neutrophils were defined as SSC-Ahigh CD66b+, CD3-, CD14-. A phagocytic score was determined using the percentage of Alexa Fluor 488+ cells and the MFI of the Alexa Fluor 488+ cells. Z-score values were calculated as described above. The previously identified mAb ZIKV-117 was used as a positive control, and mAb FLU-5J8 was used as a negative control.
Gilchuk P., Bombardi R.G., Erasmus J.H., Tan Q., Nargi R., Soto C., Abbink P., Suscovich T.J., Durnell L.A., Khandhar A., Archer J., Liang J., Fouch M.E., Davidson E., Doranz B.J., Jones T., Larson E., Ertel S., Granger B., Fuerte-Stone J., Roy V., Broge T., Linnekin T.C., Linde C.H., Gorman M.J., Nkolola J., Alter G., Reed S.G., Barouch D.H., Diamond M.S., Crowe JE J.r., Van Hoeven N., Thackray L, & Carnahan R. (2020). Integrated pipeline for the accelerated discovery of antiviral antibody therapeutics. Nature biomedical engineering, 4(11), 1030-1043.
+ Open protocol
+ Expand
8

Quantifying Antibody-Mediated NK Cell Activation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Recombinant EBOV GP ΔTM (IBT Bioservices) was coated onto a MaxiSorp 96 well plates (Nunc) at 300 ng/well at 4°C for 18 hrs. Wells were washed three times with PBS and blocked with 5% BSA in PBS. Antibodies were diluted to 10 μg/mL in PBS, and added to the plates, and were incubated for an additional 2 hrs at 37°C. Unbound antibodies were removed by washing three times with PBS, and human NK cells freshly isolated from peripheral blood of human donors by negative selection (Stem Cell Technologies, Canada) were added at 5 × 104 cells/well in the presence of 4 μg/mL brefeldin A (Sigma-Aldrich Aldrich) and 5 μg/mL GolgiStop (Life Technologies) and anti-CD107a antibody (PE-Cy5, Clone H4A3, BD Biosciences). Plates were incubated for 5 hrs at 37°C. Cells were stained for NK cell markers (CD56 PE-Cy7, clone B159, BD Biosciences; CD16 APC-Cy7, clone 3G8, BD Biosciences; CD3 AF700, clone UCHT1, BD Biosciences), followed by fixation and permeabilization with Fix and Perm (Life Technologies) according to the manufacturer’s instructions to stain for intracellular IFNγ (APC, Clone B27, BD Biosciences) and MIP-1β (PE, Clone D21–1351, BD Biosciences). Cells were analyzed on a BD LSRII flow cytometer. The glycan cap-specific mAb c13C6 (IBT Bioservices) was used as a positive control, and the DENV-specific mAb 2D22 was used as a negative control.
Gilchuk P., Murin C.D., Milligan J.C., Cross R.W., Mire C.E., Ilinykh P.A., Huang K., Kuzmina N., Altman P.X., Hui S., Gunn B.M., Bryan A.L., Davidson E., Doranz B.J., Turner H.L., Alkutkar T., Flinko R., Orlandi C., Carnahan R., Nargi R., Bombardi R.G., Vodzak M.E., Li S., Okoli A., Ibeawuchi M., Ohiaeri B., Lewis G.K., Alter G., Bukreyev A., Saphire E.O., Geisbert T.W., Ward A.B, & Crowe JE J.r. (2020). Analysis of a Therapeutic Antibody Cocktail Reveals Determinants for Cooperative and Broad Ebolavirus Neutralization. Immunity, 52(2), 388-403.e12.
+ Open protocol
+ Expand
9

Antibody-Mediated NK Cell Activation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Recombinant EBOV GP ΔTM (IBT Bioservices) was coated onto a MaxiSorp 96 well plates (Nunc) at 300 ng/well at 4°C for 18 h. Wells were washed three times with PBS and blocked with 5% BSA in PBS. Antibodies were diluted to 10 μg/mL in PBS, and added to the plates, and were incubated for an additional 2 h at 37°C. Unbound antibodies were removed by washing three times with PBS, and human NK cells freshly isolated from peripheral blood of human donors by negative selection (Stem Cell Technologies, Canada) were added at 5 Ч 104 cells/well in the presence of 4 μg/mL brefeldin A (Sigma-Aldrich Aldrich) and 5 μg/mL GolgiStop (Life Technologies) and anti-CD107a antibody (PE-Cy5, Clone H4A3, BD Biosciences). Plates were incubated for 5 h at 37°C. Cells were stained for NK cell markers (CD56 PE-Cy7, clone B159, BD Biosciences; CD16 APC-Cy7, clone 3G8, BD Biosciences; CD3 AF700, clone UCHT1, BD Biosciences), followed by fixation and permeabilization with Fix and Perm (Life Technologies) according to the manufacturer’s instructions to stain for intracellular IFNγ (APC, Clone B27, BD Biosciences) and MIP-1β (PE, Clone D21-1351, BD Biosciences). Cells were analyzed on a BD LSRII flow cytometer. The glycan cap-specific mAb c13C6 (IBT Bioservices) was used as a positive control, and the DENV-specific mAb 2D22 was used as a negative control.
Gilchuk P., Murin C.D., Milligan J.C., Cross R.W., Mire C.E., Ilinykh P.A., Huang K., Kuzmina N., Altman P.X., Hui S., Gunn B.M., Bryan A.L., Davidson E., Doranz B.J., Turner H.L., Alkutkar T., Flinko R., Orlandi C., Carnahan R., Nargi R., Bombardi R.G., Vodzak M.E., Li S., Okoli A., Ibeawuchi M., Ohiaeri B., Lewis G.K., Alter G., Bukreyev A., Saphire E.O., Geisbert T.W., Ward A.B, & Crowe JE J.r. (2020). Analysis of a Therapeutic Antibody Cocktail Reveals Determinants for Cooperative and Broad Ebolavirus Neutralization. Immunity, 52(2), 388-403.e12.
+ 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!