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SP140 Knockout Z-138 Cell Line

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LM02170079263

Product ID: LM02170079263

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隐藏域元素占位

  • 产品描述
  • 细胞复苏
  • 细胞传代
  • 细胞冻存
  • 抗体验证结果
    • Commodity name: SP140 Knockout Z-138 Cell Line
    • Commodity ID: LM02170079263
    • Gene Symbol: SP140 LYSP100
    • Ensembl ID: ENSG00000079263
    • Uniprot ID: Q13342
    • 宿主细胞 / 类型: Human mantle cell lymphoma cells
    • NCBI Gene ID: 11262
    • 规格: 1×10^6 cells/frozen vial
    • 生长培养基: Specialized culture medium
    • 筛选标记: N/A
    • 生长特性: Suspension cells, lymphoblastoid
    • 培养条件: Incubator at 37°C with 5% CO2; passage at 1/2 to 1/4 confluence.
    • 倍增时间: ~24-48 hours
    • 参考换液频率: 2–3 times per week
    • 支原体检测结果: Negative
    • 敲除效率(Sanger测序): 100%
    • 蛋白质组验证结果: Protein-level validation has been completed.
    • 抗体货号: Adding...
    • 目标基因介绍: Component of the nuclear body, also known as nuclear domain 10, PML oncogenic domain, and KR body (PubMed:8910577). May be involved in the pathogenesis of acute promyelocytic leukemia and viral infection (PubMed:8910577). May play a role in chromatin-mediated regulation of gene expression although it does not bind to histone H3 tails (PubMed:24267382).
    • 细胞开发路径: A stable KO cell line was generated using the CRISPR‑RNP approach; Sanger sequencing confirmed 100% knockout efficiency in the KO cell line.
    • 应用: A gene knockout cell line with high knockout efficiency, particularly well suited for preliminary functional analyses, the development of complex disease models, precision drug screening, and broad‑scale gene discovery studies.
  • 01. Preheat the complete culture medium in a 37°C water bath.
    02. Thaw the cryovial in a 37°C water bath for 1–2 minutes.
    03. Transfer the cryovials to a biosafety cabinet and wipe their surfaces with 70% ethanol.
    04. Unscrew the cap of the cryovial and gently transfer the cell suspension into a sterile centrifuge tube containing 9 mL of complete culture medium.
    05. Centrifuge at room temperature at 125g for 5–7 minutes, then discard the supernatant.
    06. Resuspend the cell pellet in 5 mL of complete culture medium, and transfer the cell suspension to a T25 culture flask.
    07. Transfer the cells to a 37°C incubator with 5% CO2 for culture.
    08. Recommended passage ratio: 1/2 to 1/4; cells reach confluence in 2–3 days.

  • 01. When the cell confluence in the culture flask reaches 80%–90% or higher, cells can be passaged.
    02. Remove the culture medium, PBS, and trypsin (0.25% Trypsin‑EDTA, Gibco, 25200‑056) from the 4°C refrigerator, place them in a 37°C water bath, and once their temperature approaches 37°C, take them out. Spray the exterior of the bottles with 75% ethanol, then transfer them to a biosafety cabinet.

    03. Remove the culture flask to be passaged from the incubator, spray the exterior of the flask with 75% ethanol, and place it inside a biosafety cabinet.
    04. To avoid dispersing the cells, gently rinse the cells along the inner wall of the culture flask with PBS; discard the wash solution, then add 2 mL of PBS to the T25 flask.
    05. Add the appropriate volume of trypsin (1.5 mL for a T75 flask, 0.5 mL for a T25 flask), and gently swirl the flask to ensure the enzyme evenly covers the cell‑covered bottom. Adjust the volume as needed based on the specific situation. After approximately 1–2 minutes, when most of the cells have detached, add the corresponding volume of complete culture medium to stop the digestion, then use a 5‑mL pipette to gently pipette up and down until all cells are fully detached.
    06. Transfer the cell suspension to a 15 mL centrifuge tube, centrifuge at 300 × g for 5 minutes, and discard the supernatant.
    07. Transfer 5 mL of complete culture medium to resuspend the cells, adjust the seeding ratio as needed, and replenish the culture flask with complete medium—add up to 13–15 mL for a T75 flask and 5 mL for a T25 flask—then supplement with 1% double antibiotic.
    08. After capping the bottle and tightening the lid, gently shake the vial to ensure uniform mixing of the cells, then place it in a 37°C incubator with 5% CO2.

  • 01. Prepare the cryopreservation solution and pre-cool it in advance.
    02. Ensure that the cells to be cryopreserved meet the cryopreservation requirements, and verify the following conditions under a microscope: healthy appearance and morphological characteristics, being in the late logarithmic growth phase, and showing no signs of contamination or senescence.
    03. Digest and centrifuge the cells (refer to the subculture protocol for specific steps).
    04. Resuspend the cells in cryopreservation medium at 1 mL per tube, gently pipette up and down to ensure uniformity, then aliquot into cryovials.
    05. Place the cells in a programmed cooling box and freeze them in a −80°C freezer.
    06. Subsequently, the cells are transferred to a liquid nitrogen tank for long-term storage.

  • In antibody validation

Classification: Gene-knockout cell line (protein levels validated)

Cell Line Information

Gene Symbol

SP140 LYSP100

NCBI Gene ID

11262

Ensembl ID

ENSG00000079263

Uniprot ID

Q13342

Screening marker

N/A

Host cell/type

Human mantle cell lymphoma cells

Specifications

1×10^6 cells/frozen vial

Growth Medium

Specialized culture medium

growth characteristics

Suspension cells, lymphoblastoid

culture condition

Incubator at 37°C with 5% CO2; passage at 1/2 to 1/4 confluence.

doubling time

~24-48 hours

Reference fluid change frequency

2–3 times per week

Mycoplasma test results

Negative

Knock-out validation

Knockout efficiency (Sanger sequencing)

100%

Proteome Validation Results

Protein-level validation has been completed.

Antibody number

Adding...

Antibody validation results

In antibody validation

Cell Line Description

Introduction of target gene

Component of the nuclear body, also known as nuclear domain 10, PML oncogenic domain, and KR body (PubMed:8910577). May be involved in the pathogenesis of acute promyelocytic leukemia and viral infection (PubMed:8910577). May play a role in chromatin-mediated regulation of gene expression although it does not bind to histone H3 tails (PubMed:24267382).

Cell development path

A stable KO cell line was generated using the CRISPR‑RNP approach; Sanger sequencing confirmed 100% knockout efficiency in the KO cell line.

Application

A gene knockout cell line with high knockout efficiency, particularly well suited for preliminary functional analyses, the development of complex disease models, precision drug screening, and broad‑scale gene discovery studies.

Cell Culture Instructions

Cell Resuscitation

01. Preheat the complete culture medium in a 37°C water bath.
02. Thaw the cryovial in a 37°C water bath for 1–2 minutes.
03. Transfer the cryovials to a biosafety cabinet and wipe their surfaces with 70% ethanol.
04. Unscrew the cap of the cryovial and gently transfer the cell suspension into a sterile centrifuge tube containing 9 mL of complete culture medium.
05. Centrifuge at room temperature at 125g for 5–7 minutes, then discard the supernatant.
06. Resuspend the cell pellet in 5 mL of complete culture medium, and transfer the cell suspension to a T25 culture flask.
07. Transfer the cells to a 37°C incubator with 5% CO2 for culture.
08. Recommended passage ratio: 1/2 to 1/4; cells reach confluence in 2–3 days.

cell passage

01. When the cell confluence in the culture flask reaches 80%–90% or higher, cells can be passaged.
02. Remove the culture medium, PBS, and trypsin (0.25% Trypsin‑EDTA, Gibco, 25200‑056) from the 4°C refrigerator, place them in a 37°C water bath, and once their temperature approaches 37°C, take them out. Spray the exterior of the bottles with 75% ethanol, then transfer them to a biosafety cabinet.

03. Remove the culture flask to be passaged from the incubator, spray the exterior of the flask with 75% ethanol, and place it inside a biosafety cabinet.
04. To avoid dispersing the cells, gently rinse the cells along the inner wall of the culture flask with PBS; discard the wash solution, then add 2 mL of PBS to the T25 flask.
05. Add the appropriate volume of trypsin (1.5 mL for a T75 flask, 0.5 mL for a T25 flask), and gently swirl the flask to ensure the enzyme evenly covers the cell‑covered bottom. Adjust the volume as needed based on the specific situation. After approximately 1–2 minutes, when most of the cells have detached, add the corresponding volume of complete culture medium to stop the digestion, then use a 5‑mL pipette to gently pipette up and down until all cells are fully detached.
06. Transfer the cell suspension to a 15 mL centrifuge tube, centrifuge at 300 × g for 5 minutes, and discard the supernatant.
07. Transfer 5 mL of complete culture medium to resuspend the cells, adjust the seeding ratio as needed, and replenish the culture flask with complete medium—add up to 13–15 mL for a T75 flask and 5 mL for a T25 flask—then supplement with 1% double antibiotic.
08. After capping the bottle and tightening the lid, gently shake the vial to ensure uniform mixing of the cells, then place it in a 37°C incubator with 5% CO2.

cell cryopreservation

01. Prepare the cryopreservation solution and pre-cool it in advance.
02. Ensure that the cells to be cryopreserved meet the cryopreservation requirements, and verify the following conditions under a microscope: healthy appearance and morphological characteristics, being in the late logarithmic growth phase, and showing no signs of contamination or senescence.
03. Digest and centrifuge the cells (refer to the subculture protocol for specific steps).
04. Resuspend the cells in cryopreservation medium at 1 mL per tube, gently pipette up and down to ensure uniformity, then aliquot into cryovials.
05. Place the cells in a programmed cooling box and freeze them in a −80°C freezer.
06. Subsequently, the cells are transferred to a liquid nitrogen tank for long-term storage.