For adoptive transfer of BM MKs, GFP mice (donor) and Mplhlb219 mice (recipient) were simultaneously exposed to a single dose of 5 Gy TBI. Then, 3 × 105 BM MKs from GFP mice, Igf1f/f or Igf1ΔMK mice at 2 dpi were obtained by continuously enriching CD41+ cells and CD42d+ cells using an EasySep™ Release Mouse Biotin Positive Selection Kit and an EasySep™ Release Mouse PE Positive Selection Kit (all StemCell Technologies, Vancouver, BC, Canada) with biotin-labeled anti-CD41 and PE-labeled anti-CD42d antibodies according to the manufacturer’s instructions. Subsequently, the freshly-isolated MKs were intravenously transfused into Mplhlb219 mice.
Easysep release mouse biotin positive selection kit
Manufactured by STEMCELL
Sourced in Canada
The EasySep™ Release Mouse Biotin Positive Selection Kit is a laboratory tool designed for the isolation and enrichment of biotin-labeled mouse cells from a sample. The kit utilizes magnetic particles to capture the target cells, allowing for their separation from the rest of the cell population.
Automatically generated - may contain errors
3 protocols using easysep release mouse biotin positive selection kit
1
Adoptive Transfer of BM MKs
2
Isolation and Analysis of Tumor-Infiltrating Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tumors were resected and weighed before being placed into RPMI containing Liberase TL Research Grade (250 µg/mL; Roche) and DNase I (1 mg/mL; Sigma-Aldrich). Tumors were minced with scissors and incubated at 37°C for 35 min for enzymatic digestion. Tumor pieces were subsequently crushed against a 70 µm filter to generate a single cell suspension. Cells were washed with PBS containing 2% FBS and 1 mM EDTA. Leucocytes were isolated from each respective single cell suspension using a EasySep Release Mouse Biotin Positive Selection Kit (Stem Cell Technologies) in conjunction with an anti-CD45.2-Biotin (Clone 104) according to the manufacturer’s guidelines. Tumor draining lymph nodes were harvested and pressed against a 70 µm filter and rinsed with Hanks Balanced Salt Solution. Blood was collected from the retro-orbital sinus and red blood cells were lysed using ACK buffer (150 mM NH4Cl, 10 mM KHCO3, and 0.1 mM Na2EDTA, pH 7.3).
3
Isolation and Enrichment of Mouse Megakaryocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bone marrow cells (BMCs) were flushed from the femur and tibia, and red blood cells were lysed using a red cell lysis buffer. Mouse mMKs were obtained by continuously enriching CD41+ cells and CD42d+ cells using an EasySep™ Release Mouse Biotin Positive Selection Kit and an EasySep™ Release Mouse Phycoerythrin Positive Selection Kit (all StemCell Technologies) with biotin-labeled anti-CD41 and Phycoerythrin-labeled anti-CD42d antibodies according to the manufacturer’s instructions. The antibodies used were listed in Additional file 1 : Table S1.
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
Revolutionizing how scientists
search and build protocols!