Log In Sign up
The largest database of trusted experimental protocols

Easysep cd8 t cell isolation kit

Manufactured by STEMCELL
Visit Supplier
Sourced in Canada

The EasySep CD8+ T Cell Isolation Kit is a laboratory product designed to isolate CD8+ T cells from a variety of sample types. The kit utilizes magnetic particles and a specialized buffer system to select and enrich for the target cell population.

Automatically generated - may contain errors

Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (4 mentions) Histopaque 1083 gradient, by Merck Group (2 mentions) Ack lysis buffer, by Thermo Fisher Scientific (2 mentions) Hepes, by Thermo Fisher Scientific (2 mentions) L glutamine, by Merck Group (2 mentions) β mercaptoethanol, by Merck Group (2 mentions) Rpmi 1640, by Corning (2 mentions) Facsaria 2, by BD (2 mentions) Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions) Live dead aqua fixable cell stain, by Thermo Fisher Scientific (1 mentions) α cd28, by Thermo Fisher Scientific (1 mentions) Interleukin 2 (il 2), by Thermo Fisher Scientific (1 mentions) α cd3, by Thermo Fisher Scientific (1 mentions) Percoll, by GE Healthcare (1 mentions) Rat igg2b, by BioXCell (1 mentions) Foxp3 transcription factor buffer set, by Thermo Fisher Scientific (1 mentions) Ack buffer, by Thermo Fisher Scientific (1 mentions) Facsaria, by BD (1 mentions) Easysep b cell isolation kit, by STEMCELL (1 mentions)

Close spelling variants of this product

EasySep™ Mouse CD8+ or CD4+ T Cell Isolation Kits EasySep Human Naive CD4+ or CD8+ T cell isolation kits EasySep cell isolation kit EasySep isolation kits Cell isolation kits EasySep kits Isolation kits

10 protocols using easysep cd8 t cell isolation kit

1

Isolation of Murine Splenocytes and PBMC

Check if the same lab product or an alternative is used in the 5 most similar protocols
Single-cell suspensions of splenocytes were generated by mechanically homogenizing spleens using a 70μm cell strainer. Cells were washed and red blood cells lysed using ACK lysis buffer (ThermoFisher). PBMCs were isolated using Histopaque®-1083 gradient (Sigma-Aldrich). When necessary, cells were sorted with a FACS Aria II (BD Biosciences) to >92% purity. Before sorting, CD8 T cells were enriched from total splenocytes using the EasySeptm CD8 T cell isolation Kit (StemCell) (routinely >85% purity). CD8 T cell-enrichment was also performed before adoptively transferring P14 cells to recipient mice. All in vitro T cell assays were performed using complete RPMI medium (cRPMI): RPMI-1640 (Corning/Mediatech), 10% fetal bovine serum (FBS) (ThermoFisher), 1% HEPES (ThermoFisher), 1% penicillin/streptomycin (ThermoFisher), 1% L-glutamine, and 0.1% β-mercaptoethanol (Sigma-Aldrich).
Abdel-Hakeem M.S., Manne S., Beltra J.C., Stelekati E., Chen Z., Nzingha K., Ali M.A., Johnson J.L., Giles J.R., Mathew D., Greenplate A.R., Vahedi G, & Wherry E.J. (2021). Epigenetic scarring of exhausted T cells hinders memory differentiation upon eliminating chronic antigenic stimulation. Nature immunology, 22(8), 1008-1019.
+ Open protocol
+ Expand
2

Tex Subsets Adoptive Transfer in LCMV

Check if the same lab product or an alternative is used in the 5 most similar protocols
C57BL/6 mice (CD45.2+) were infected with LCMV clone 13 and spleens were collected at day 21 pi. Mice with low viral titers in the serum at day 15pi were excluded. CD8+ T cells were enriched using EasySeptm CD8+ T cell isolation Kit (StemCell) (routinely > 90% purity) and Tex subsets were sorted from endogenous activated CD8+ T cells. Briefly, Live/Dead Fixable Aqua Cell Stain (ThermoFisher Scientific), CD4 and CD19 were used as exclusion markers and PD-1, Ly108 and CD69 were used to discriminate and sort Tex subsets among endogenous activated (PD-1+) CD8+ T cells (purity was routinely > 94% for each subset). Sorted cells were stained with Carboxyfluorescein succinimidyl ester (CFSE; ThermoFisher Scientific) for 8min at RT in PBS containing the CFSE dye (5μM). Reaction was stopped by adding an equal volume of cold FBS and cells were subsequently washed two times in cRPMI. Cells were counted and 1x105 of each subset was adoptively transferred into infection-matched CD45.1+ recipient mice. Proliferation and phenotypic changes were assessed in the spleen seven days post-transfer.
Beltra J.C., Manne S., Abdel-Hakeem M.S., Kurachi M., Giles J.R., Chen Z., Casella V., Ngiow S.F., Khan O., Huang Y.J., Yan P., Nzingha K., Xu W., Amaravadi R.K., Xu X., Karakousis G.C., Mitchell T.C., Schuchter L.M., Huang A.C, & Wherry E.J. (2020). Developmental Relationships of Four Exhausted CD8+ T Cell Subsets Reveals Underlying Transcriptional and Epigenetic Landscape Control Mechanisms. Immunity, 52(5), 825-841.e8.
+ Open protocol
+ Expand
3

Transduction of P14 CD8+ T cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
RV transduction of P14 CD8+ T cells was performed as described (Kurachi et al., 2017 (link)). P14 CD8+ T cells were enriched from total splenocytes using EasySeptm CD8+ T cell isolation Kit (StemCell) and activated in vitro in cRPMI supplemented with αCD3 (1 μg/mL), αCD28 (0.5μg/mL) antibodies and IL-2 (100U/mL) (PeproTech). One day post activation (between 24-27 h), activated CD8+ T cells were enriched using Percoll (GE Healthcare) density gradient (30%/60%) and spin-transduced during 60-75min at 2000g 30°C with RV supernatant containing polybrene (4μg/mL). Transduced cells were then incubated for 6 h, washed twice in cRPMI, counted and injected (1x105 per mouse) into LCMV clone 13 infected mice at day 1.5pi.
Beltra J.C., Manne S., Abdel-Hakeem M.S., Kurachi M., Giles J.R., Chen Z., Casella V., Ngiow S.F., Khan O., Huang Y.J., Yan P., Nzingha K., Xu W., Amaravadi R.K., Xu X., Karakousis G.C., Mitchell T.C., Schuchter L.M., Huang A.C, & Wherry E.J. (2020). Developmental Relationships of Four Exhausted CD8+ T Cell Subsets Reveals Underlying Transcriptional and Epigenetic Landscape Control Mechanisms. Immunity, 52(5), 825-841.e8.
+ Open protocol
+ Expand
4

Isolation of Murine Splenocytes and PBMC

Check if the same lab product or an alternative is used in the 5 most similar protocols
Single-cell suspensions of splenocytes were generated by mechanically homogenizing spleens using a 70μm cell strainer. Cells were washed and red blood cells lysed using ACK lysis buffer (ThermoFisher). PBMCs were isolated using Histopaque®-1083 gradient (Sigma-Aldrich). When necessary, cells were sorted with a FACS Aria II (BD Biosciences) to >92% purity. Before sorting, CD8 T cells were enriched from total splenocytes using the EasySeptm CD8 T cell isolation Kit (StemCell) (routinely >85% purity). CD8 T cell-enrichment was also performed before adoptively transferring P14 cells to recipient mice. All in vitro T cell assays were performed using complete RPMI medium (cRPMI): RPMI-1640 (Corning/Mediatech), 10% fetal bovine serum (FBS) (ThermoFisher), 1% HEPES (ThermoFisher), 1% penicillin/streptomycin (ThermoFisher), 1% L-glutamine, and 0.1% β-mercaptoethanol (Sigma-Aldrich).
Abdel-Hakeem M.S., Manne S., Beltra J.C., Stelekati E., Chen Z., Nzingha K., Ali M.A., Johnson J.L., Giles J.R., Mathew D., Greenplate A.R., Vahedi G, & Wherry E.J. (2021). Epigenetic scarring of exhausted T cells hinders memory differentiation upon eliminating chronic antigenic stimulation. Nature immunology, 22(8), 1008-1019.
+ Open protocol
+ Expand
5

ZIKV Infection Model with CD8+ T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Four- to 5-week-old male and female HLA-B*0702 Ifnar1−/− mice were vaccinated and boosted with NS3 vaccine or saline as described above. Spleens were harvested on day 49, and CD8+ T cells were purified from the spleens by negative selection using the EasySep CD8+ T Cell Isolation Kit (StemCell). Purified CD8+ T cells (107 per mouse) were intravenously injected into 7- to 8-week-old HLA-B*0702 Ifnar1−/− recipient mice. One day later, the recipient mice were infected retro-orbitally with ZIKV SD001. For the depletion experiments, vaccinated and boosted mice were intravenously injected with 300 μg of a CD8-depleting monoclonal antibody (mAb) (2.43, Bio X Cell) or an isotype control mAb (rat IgG2b, Bio X Cell) on days −3 and −1 before infection retro-orbitally with ZIKV SD001. In both sets of experiments, blood and organs were harvested 3 days later and ZIKV RNA was quantified.
Elong Ngono A., Syed T., Nguyen A.V., Regla-Nava J.A., Susantono M., Spasova D., Aguilar A., West M., Sparks J., Gonzalez A., Branche E., DeHart J.L., Vega J.B., Karmali P.P., Chivukula P., Kamrud K., Aliahmad P., Wang N, & Shresta S. (2020). CD8+ T cells mediate protection against Zika virus induced by an NS3-based vaccine. Science Advances, 6(45), eabb2154.
+ Open protocol
+ Expand
6

Quantifying Intracellular TGFβ Inhibitor Delivery

Check if the same lab product or an alternative is used in the 5 most similar protocols
Primary CD8+ T cells were isolated from spleens of C57Bl/6 mice using an EasySep™ CD8+ T cell isolation kit (Stemcell Technologies). T cells were incubated in complete medium (RPMI containing 10% fetal bovine serum) for 4 h in the presence of amph-NPs loaded with TGF-βi or free TGF-βi. The concentration of drug per cell was then determined by HPLC following cell membrane permeabilization and small molecule extraction in ethanol. To rupture cell membranes, 106 CD8+ T cells were sonicated in a water bath for two minutes at 25 °C. To extract cytosolic TGFβi into the supernatant, 190 uL of ethanol and 10 uL of β-mercapoethanol were added to the cell suspension. The mixture was then placed on a shaker overnight at 25°C. The resulting cell lysates were centrifuged at 14 Kxg for 10 minutes, and the supernatant containing solubilized TGFbi was loaded into HPLC test vials (80 uL per sample was injected). Samples were run through a reversed phase C18 column (Gemini® 5 µm C18 110 Å, LC Column 250 × 4.6 mm), with a 30 min protocol 20% – 95% (Acetonitrile + 0.1% TFA; water + 0.1% TFA). Recovered TGFβi was detected at ~12.5 min elution time by UV spectroscopy at 350 nm.
Yang Y.S., Moynihan K.D., Bekdemir A., Dichwalkar T.M., Noh M.M., Watson N., Melo M., Ingram J., Suh H., Ploegh H., Stellacci F.R, & Irvine D.J. (2018). Targeting small molecule drugs to T cells with antibody-directed cell-penetrating gold nanoparticles. Biomaterials science, 7(1), 113-124.
+ Open protocol
+ Expand
7

CD8+ T Cell Isolation and Sorting

Check if the same lab product or an alternative is used in the 5 most similar protocols
Spleens were mechanically disrupted onto a 70-μM cell strainer and red blood cells were lysed with ACK buffer (Gibco). Cells were stained with extracellular antibodies for 30 min on ice. For transcription-factor detection, cells were fixed and permeabilized using the Foxp3 Transcription Factor buffer set (Thermo Fisher Scientific). Samples were acquired on an LSR II and analyzed with FlowJo, version 10 software (Tree Star Inc). For cell sorting, CD8+ T cells were enriched using the EasySep CD8+ T Cell Isolation Kit (StemCell) and VEX+ cells were sorted based on CD8, CD45.1, CD45.2, and VEX on a BD FACSARIA (BD Bioscience) using a 70-μm nozzle.
Stelekati E., Cai Z., Manne S., Chen Z., Beltra J.C., Buchness L.A., Leng X., Ristin S., Nzingha K., Ekshyyan V., Niavi C., Abdel-Hakeem M.S., Ali M.A., Drury S., Lau C.W., Gao Z., Ban Y., Zhou S.K., Ansel K.M., Kurachi M., Jordan M.S., Villarino A.V., Ngiow S.F, & Wherry E.J. (2022). MicroRNA-29a attenuates CD8 T cell exhaustion and induces memory-like CD8 T cells during chronic infection. Proceedings of the National Academy of Sciences of the United States of America, 119(17), e2106083119.
+ Open protocol
+ Expand
8

Investigating Memory CD8+ T Cell Responses

Check if the same lab product or an alternative is used in the 5 most similar protocols
For transfer and recall experiments, CD8 T cells were enriched from splenocytes of CD45.2 mice at day 30 or ≥95 after 3° using an EasySep CD8+ T cell isolation kit (STEMCELL Technologies, Vancouver, Canada). For early memory transfers, 2.5 × 105 Kb-SIINFEKL-specific CD8 T cells were transferred i.v. to CD45.1 mice. For late memory transfer experiments, cells were flow sorted and either 2 × 103 or 2 × 105 Kb-SIINFEKL-specific CD8 T cells were transferred i.v. to CD45.1 mice. In all cases, recipient mice were infected i.v. with 1 × 106 PFU of VSV-OVA the day after cell transfer.
To determine the ability of T cells to protect upon re-infection, 3° short-boosted and long-boosted mice were generated as described above and sacrificed five days after infection with 2 × 106 PFU VV-OVA i.v. (the 3° boost). Ovaries and spleen were harvested, homogenized and viral loads were analyzed by plaque assays on 143B cells, as previously described (31 ). Naïve mice were infected with 2 × 106 PFU VV-OVA i.v. as a control.
Thompson E.A., Beura L.K., Nelson C.E., Anderson K.G, & Vezys V. (2016). Shortened intervals during heterologous boosting preserve memory CD8 T cell function but compromise longevity. Journal of immunology (Baltimore, Md. : 1950), 196(7), 3054-3063.
+ Open protocol
+ Expand
9

Isolation and Purification of Naive and Memory CD8+ T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Peripheral blood mononuclear cells (PBMCs) were isolated using a Ficoll-Hypaque density gradient and either used immediately or stored in liquid nitrogen. CD8+ T cells from normal PBMCs were negatively selected using EasySep CD8+ T-cell isolation kit (StemCell Technologies). To isolate CD8+ T cells from CLL patient samples, CD19+ B cells were first depleted using EasySep B-cell isolation kit (StemCell Technologies) before undergoing CD8+ T cells negative selection. To further enrich memory and naïve T cells from negatively selected CD8+ T cells, we used biotin-conjugated antibodies against CD45RO and CD244 (2B4), which are predominantly expressed by effector and memory T cells, for positive selection of memory T cells. The flow-through fraction is untouched naïve CD8+ T cells. Using this approach, we can obtain high purity of naïve and memory CD8+ T cells (greater than 90% purity).
Wu J., Xu X., Lee E.J., Shull A.Y., Pei L., Awan F., Wang X., Choi J.H., Deng L., Xin H.B., Zhong W., Liang J., Miao Y., Wu Y., Fan L., Li J., Xu W, & Shi H. (2016). Phenotypic alteration of CD8+ T cells in chronic lymphocytic leukemia is associated with epigenetic reprogramming. Oncotarget, 7(26), 40558-40570.
+ Open protocol
+ Expand
10

Exosome-mediated CD8+ T cell activation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Isolation of CD8+ T cells from human peripheral blood mononuclear cells (PBMCs) was performed using the Easy‐Sep™ CD8+ T cell Isolation Kit (Stemcell Technologies). Exosomes derived from TNBC cells were added into a 12‐well plate and co‐cultured with CD3/28 bead (MBS‐C001; AcroBiosystems) preactivated CD8+ T cells. Further analysis was performed after incubation for 24 h.
Li W., Han G., Li F., Bu P., Hao Y., Huang L, & Bai X. (2023). Cancer cell‐derived exosomal miR‐20a‐5p inhibits CD8 + T‐cell function and confers anti‐programmed cell death 1 therapy resistance in triple‐negative breast cancer. Cancer Science, 115(2), 347-356.
+ 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!