Ack lysing buffer

Manufactured by BD

ACK lysing buffer is a solution used for the lysis of red blood cells (erythrocytes) in biological samples. It is a common reagent used in various laboratory procedures, particularly in the preparation of cell suspensions for flow cytometry, hematological analysis, and other applications that require the separation of white blood cells (leukocytes) from red blood cells.

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Lab products found in correlation

Stemspan medium, by STEMCELL (1 mentions) Penicillin streptomycin, by Solarbio (1 mentions) M csf, by R&D Systems (1 mentions) Cd4 cd25 t cell isolation kit, by Miltenyi Biotec (1 mentions)

Close spelling variants of this product

Lysing buffer (117 citations) ACK buffer (8 citations)

3 protocols using ack lysing buffer

Culturing Bone Marrow-Derived Macrophages and Hematopoietic Stem Cells

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For bone marrow-derived macrophages (BMDMs) culturing, femurs were flushed to get bone marrow cells. The bone marrow cells were cultured with DMEM medium that containing 10% FBS, 1% penicillin–streptomycin (Solarbio, P1400) and 20 ng/ml m-CSF (R&D, 416-M-050). The medium was refreshed at day 5 and added with 20 ng/ml of IL-4 for 4 days to induce M2-like BMDMs, during which vehicle or 10 μM NE was added to the culture. For in vitro analyses of HSCs, bone marrow cells were treated with ACK lysing buffer (BD Biosciences) for 2 min to exclude red blood cells. Then the lineage-depleted bone marrow cells were obtained through Hematopoietic Cell Lineage Depletion Kit (R&D Systems, MAGM209). The lineage-depleted bone marrow cells were cultured for 7 days in StemSpan medium (StemCell Technologies) containing SCF (R&D Systems, 445-MC, 10 ng/ml), Fgf1 (R&D Systems, 4686-FA, 10 ng/ml) and Thpo (R&D Systems, 488-TO, 20 ng/ml).^{69 (link)} Vehicle or 10 μM NE was added to assess NE-related effects on HSCs.

Liu W., Chen W., Xie M., Chen C., Shao Z., Zhang Y., Zhao H., Song Q., Hu H., Xing X., Cai X., Deng X., Li X., Wang P., Liu G., Xiong L., Lv X, & Zhang Y. (2023). Traumatic brain injury stimulates sympathetic tone-mediated bone marrow myelopoiesis to favor fracture healing. Signal Transduction and Targeted Therapy, 8, 260.

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Adoptive Transfer of Mouse Tregs

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Spleens from C57BL/6 mice were harvested from animals 2 months after muscle gene transfer with AAV1-hFIX vector and from age matched naive animals into 2-MLC media (DMEM, 2% heat-inactivated fetal calf serum, 1 mM sodium pyruvate, 10 mM HEPES, 0.1 mM non-essential amino acids, 10-6 M 2-mercaptoethanol, and antibiotics) at room temperature, homogenized, filtered through a 70-µm cell strainer, and centrifuged at 300 g for 10 min. Cells were subsequently incubated with ACK lysing buffer (BD Bioscience) for 5 min and washed twice with 2-MLC medium. Viable splenocytes were counted with a hemocytometer and trypan blue. The CD4⁺CD25⁺ T cell isolation kit (Miltenyi, Auburn, CA) was used to purify Tregs from bulk splenocytes by magnetic cell sorting as described [25 (link)]. Purified CD4⁺CD25⁺ cells were pooled for each experimental group of donor mice and delivered to naive C57BL/6 mice at 1×10⁶ by tail vein injection. Recipient mice were immunized with 5µg hFIX in Complete Freund’s Adjuvant (CFA) 24 hours after adoptive transfer.

Herzog R.W., Cooper M., Perrin G.Q., Biswas M M., Martino A.T., Morel L., Terhorst C, & Hoffman B.E. (2017). Regulatory T cells and TLR9 Activation Shape Antibody Formation to a Secreted Transgene Product in AAV Muscle Gene Transfer. Cellular immunology.

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Isolation of Primary Mouse Liver Cells

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Primary liver cells were isolated by a combination of collagenase digestion and centrifugal elutriation as described (22 (link), 23 (link)). Briefly, the mouse livers were perfused through the superior vena cava by 0.2 mM EGTA and 5 mg/ml Liberase TM. The liver cell suspensions were filtered through a 100-μm cell strainer to eliminate undigested tissue remnants. Hepatocytes were collected from the pellet after centrifugation twice at 500 rpm for 3 min. The supernatant was centrifuged at 300g for 10 min. The pellet was resuspended in 35% (vol/vol) Percoll Plus solution and centrifuged at 500g for 15 min. NPCs were collected from the pellet, which was suspended in ACK Lysing Buffer from BD Biosciences and washed.

Wang J., Zhang Z., Guan J., Tung H.C., Xie J., Huang H., Chen Y., Xu M., Ren S., Li S., Zhang M., Yang D, & Xie W. (2023). Hepatocyte estrogen sulfotransferase inhibition protects female mice from concanavalin A–induced T cell–mediated hepatitis independent of estrogens. The Journal of Biological Chemistry, 299(3), 103026.

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