BSG Knockout A549 Cell Line
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Product ID： LM02100172270
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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
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- Commodity name: BSG Knockout A549 Cell Line
- Commodity ID: LM02100172270
- Gene Symbol: BSG UNQ6505/PRO21383
- Ensembl ID: ENSG00000172270
- Uniprot ID: P35613
- 宿主细胞 / 类型: A549/human non-small cell lung cancer cells
- NCBI Gene ID: 682
- 规格: 1×10^6 cells/frozen vial
- 生长培养基: MEM+10%FBS+100U,ml Penicillin-Strep
- 筛选标记: N/A
- 生长特性: Adherent cells, epithelial-like
- 培养条件: Incubate at 37°C in a 5% CO2 atmosphere, and passage at a ratio of 1/2 to 1/3.
- 倍增时间: ~22 hours
- 参考换液频率: Change the liquid every 2–3 days.
- 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: [Isoform 1]: (Microbial infection) Erythrocyte receptor for P.falciparum RH5 which is essential for erythrocyte invasion by the merozoite stage of P.falciparum isolates 3D7 and Dd2.||[Isoform 1]: Essential for normal retinal maturation and development (By similarity). Acts as a retinal cell surface receptor for NXNL1 and plays an important role in NXNL1-mediated survival of retinal cone photoreceptors (PubMed:25957687). In association with glucose transporter SLC16A1/GLUT1 and NXNL1, promotes retinal cone survival by enhancing aerobic glycolysis and accelerating the entry of glucose into photoreceptors (PubMed:25957687). May act as a potent stimulator of IL6 secretion in multiple cell lines that include monocytes (PubMed:21620857).||[Isoform 2]: (Microbial infection) Acts as a receptor for measles virus.||[Isoform 2]: (Microbial infection) Acts as a receptor for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).||[Isoform 2]: (Microbial infection) Can facilitate HIV-1 infection via its interaction with virus-associated PPIA/CYPA.||[Isoform 2]: (Microbial infection) Can facilitate human SARS coronavirus (SARS-CoV-1) infection via its interaction with virus-associated PPIA/CYPA.||[Isoform 2]: (Microbial infection) Erythrocyte receptor for P.falciparum RH5 which is essential for erythrocyte invasion by the merozoite stage of P.falciparum isolates 3D7, Dd2, 7G8 and HB3 (PubMed:22080952, PubMed:26195724). Binding of P.falciparum RH5 results in BSG dimerization which triggers an increase in intracellular Ca(2+) in the erythrocyte (PubMed:28409866). This essential step leads to a rearrangement of the erythrocyte cytoskeleton required for the merozoite invasion (PubMed:28409866).||[Isoform 2]: (Microbial infection) Promotes entry of pentamer-expressing human cytomegalovirus (HCMV) into epithelial and endothelial cells.||[Isoform 2]: Signaling receptor for cyclophilins, essential for PPIA/CYPA and PPIB/CYPB-dependent signaling related to chemotaxis and adhesion of immune cells (PubMed:11943775, PubMed:11688976). Plays an important role in targeting monocarboxylate transporters SLC16A1/GLUT1, SLC16A11 and SLC16A12 to the plasma membrane (PubMed:17127621, PubMed:21778275, PubMed:28666119). Acts as a coreceptor for vascular endothelial growth factor receptor 2 (KDR/VEGFR2) in endothelial cells enhancing its VEGFA-mediated activation and downstream signaling (PubMed:25825981). Promotes angiogenesis through EPAS1/HIF2A-mediated up-regulation of VEGFA (isoform VEGF-165 and VEGF-121) and KDR/VEGFR2 in endothelial cells (PubMed:19837976). Plays a key role in regulating tumor growth, invasion, metastasis and neoangiogenesis by stimulating the production and release of extracellular matrix metalloproteinases and KDR/VEGFR2 by both tumor cells and stromal cells (fibroblasts and endothelial cells) (PubMed:12553375, PubMed:11992541, PubMed:15833850).
- 细胞开发路径: A stable KO cell line was generated using the CRISPR-RNP method; Sanger sequencing confirmed 100% knockout efficiency in the KO cell line.
- 应用: Gene knockout cell lines (KO Cell Lines) with high knockout efficiency are particularly well suited for preliminary functional analyses, the development of complex disease models, precision drug screening, and broad gene discovery studies.
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01. Pre-warm 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 cryovial to a biosafety cabinet and wipe its surface 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 125g at room temperature 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, 5% CO2 incubator for culture.
08. Recommended passage ratio: Passage at a ratio of 1/2 to 1/3; cells will confluence in 2–3 days. -
01. Cell passage can be performed when the cell confluence in the culture flask reaches 80%–90% or higher.
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 remove them when their temperature approaches 37°C. After spraying the surfaces of the bottles with 75% ethanol, transfer them to a biosafety cabinet.03. Remove the culture flask to be passaged from the incubator, spray the flask with 75% ethanol, and place it in a biosafety cabinet.
04. To avoid detaching the cells, rinse them with PBS along the inner wall of the culture flask; discard the wash solution, and add 2 mL to a 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 so the enzyme evenly covers the bottom. Adjust the volume as necessary. After about 1–2 minutes, when most cells have detached, add the corresponding volume of complete culture medium to neutralize the trypsin, then gently pipette up and down with a 5 mL pipette until all cells are fully detached.
06. Transfer the cell suspension to a 15 mL centrifuge tube, centrifuge at 300g for 5 min, and discard the supernatant.
07. Resuspend the cells in 5 mL of complete medium, adjust the seeding ratio as needed, and replenish the complete medium in the culture flasks—add to 13–15 mL for T75 flasks and to 5 mL for T25 flasks—and supplement with 1% double antibiotic.
08. After capping the bottle and tightening the cap, gently shake the bottle to mix the cells evenly, then place it in a 37°C, 5% CO2 incubator. -
01. Prepare the cryopreservation solution and pre-cool it in advance.
02. Ensure that the cells to be cryopreserved meet the cryopreservation requirements; examine the following conditions under a microscope: healthy appearance and morphological characteristics, appropriate growth phase (late logarithmic phase), and absence of contamination or signs of decline.
03. Digest and centrifuge the cells (refer to the subculture protocol for specific steps).
04. Resuspend the cells in cryopreservation solution at 1 mL per tube, mix thoroughly by pipetting, and aliquot into cryovials.
05. Place the cells in a programmed cooling box and freeze them in a -80°C freezer.
06. The cells are subsequently transferred to a liquid nitrogen tank for long-term storage. - Western blot (WB) analysis was performed simultaneously on lysates from wild-type and KO cells using a specific monoclonal antibody; the target protein band was detected in WT cells but absent or diminished in KO cells.
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Classification： Gene-knockout cell line (protein levels validated)
Cell Line Information
Gene Symbol
BSG UNQ6505/PRO21383
NCBI Gene ID
682
Ensembl ID
ENSG00000172270
Uniprot ID
P35613
Screening marker
N/A
Host cell/type
A549/human non-small cell lung cancer cells
Specifications
1×10^6 cells/frozen vial
Growth Medium
MEM+10%FBS+100U,ml Penicillin-Strep
growth characteristics
Adherent cells, epithelial-like
culture condition
Incubate at 37°C in a 5% CO2 atmosphere, and passage at a ratio of 1/2 to 1/3.
doubling time
~22 hours
Reference fluid change frequency
Change the liquid every 2–3 days.
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
Cell Line Description
Introduction of target gene
[Isoform 1]: (Microbial infection) Erythrocyte receptor for P.falciparum RH5 which is essential for erythrocyte invasion by the merozoite stage of P.falciparum isolates 3D7 and Dd2.||[Isoform 1]: Essential for normal retinal maturation and development (By similarity). Acts as a retinal cell surface receptor for NXNL1 and plays an important role in NXNL1-mediated survival of retinal cone photoreceptors (PubMed:25957687). In association with glucose transporter SLC16A1/GLUT1 and NXNL1, promotes retinal cone survival by enhancing aerobic glycolysis and accelerating the entry of glucose into photoreceptors (PubMed:25957687). May act as a potent stimulator of IL6 secretion in multiple cell lines that include monocytes (PubMed:21620857).||[Isoform 2]: (Microbial infection) Acts as a receptor for measles virus.||[Isoform 2]: (Microbial infection) Acts as a receptor for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).||[Isoform 2]: (Microbial infection) Can facilitate HIV-1 infection via its interaction with virus-associated PPIA/CYPA.||[Isoform 2]: (Microbial infection) Can facilitate human SARS coronavirus (SARS-CoV-1) infection via its interaction with virus-associated PPIA/CYPA.||[Isoform 2]: (Microbial infection) Erythrocyte receptor for P.falciparum RH5 which is essential for erythrocyte invasion by the merozoite stage of P.falciparum isolates 3D7, Dd2, 7G8 and HB3 (PubMed:22080952, PubMed:26195724). Binding of P.falciparum RH5 results in BSG dimerization which triggers an increase in intracellular Ca(2+) in the erythrocyte (PubMed:28409866). This essential step leads to a rearrangement of the erythrocyte cytoskeleton required for the merozoite invasion (PubMed:28409866).||[Isoform 2]: (Microbial infection) Promotes entry of pentamer-expressing human cytomegalovirus (HCMV) into epithelial and endothelial cells.||[Isoform 2]: Signaling receptor for cyclophilins, essential for PPIA/CYPA and PPIB/CYPB-dependent signaling related to chemotaxis and adhesion of immune cells (PubMed:11943775, PubMed:11688976). Plays an important role in targeting monocarboxylate transporters SLC16A1/GLUT1, SLC16A11 and SLC16A12 to the plasma membrane (PubMed:17127621, PubMed:21778275, PubMed:28666119). Acts as a coreceptor for vascular endothelial growth factor receptor 2 (KDR/VEGFR2) in endothelial cells enhancing its VEGFA-mediated activation and downstream signaling (PubMed:25825981). Promotes angiogenesis through EPAS1/HIF2A-mediated up-regulation of VEGFA (isoform VEGF-165 and VEGF-121) and KDR/VEGFR2 in endothelial cells (PubMed:19837976). Plays a key role in regulating tumor growth, invasion, metastasis and neoangiogenesis by stimulating the production and release of extracellular matrix metalloproteinases and KDR/VEGFR2 by both tumor cells and stromal cells (fibroblasts and endothelial cells) (PubMed:12553375, PubMed:11992541, PubMed:15833850).
Cell development path
A stable KO cell line was generated using the CRISPR-RNP method; Sanger sequencing confirmed 100% knockout efficiency in the KO cell line.
Application
Gene knockout cell lines (KO Cell Lines) with high knockout efficiency are particularly well suited for preliminary functional analyses, the development of complex disease models, precision drug screening, and broad gene discovery studies.
Cell Culture Instructions
Cell Resuscitation
01. Pre-warm 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 cryovial to a biosafety cabinet and wipe its surface 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 125g at room temperature 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, 5% CO2 incubator for culture.
08. Recommended passage ratio: Passage at a ratio of 1/2 to 1/3; cells will confluence in 2–3 days.
cell passage
01. Cell passage can be performed when the cell confluence in the culture flask reaches 80%–90% or higher.
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 remove them when their temperature approaches 37°C. After spraying the surfaces of the bottles with 75% ethanol, transfer them to a biosafety cabinet.
03. Remove the culture flask to be passaged from the incubator, spray the flask with 75% ethanol, and place it in a biosafety cabinet.
04. To avoid detaching the cells, rinse them with PBS along the inner wall of the culture flask; discard the wash solution, and add 2 mL to a 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 so the enzyme evenly covers the bottom. Adjust the volume as necessary. After about 1–2 minutes, when most cells have detached, add the corresponding volume of complete culture medium to neutralize the trypsin, then gently pipette up and down with a 5 mL pipette until all cells are fully detached.
06. Transfer the cell suspension to a 15 mL centrifuge tube, centrifuge at 300g for 5 min, and discard the supernatant.
07. Resuspend the cells in 5 mL of complete medium, adjust the seeding ratio as needed, and replenish the complete medium in the culture flasks—add to 13–15 mL for T75 flasks and to 5 mL for T25 flasks—and supplement with 1% double antibiotic.
08. After capping the bottle and tightening the cap, gently shake the bottle to mix the cells evenly, then place it in a 37°C, 5% CO2 incubator.
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; examine the following conditions under a microscope: healthy appearance and morphological characteristics, appropriate growth phase (late logarithmic phase), and absence of contamination or signs of decline.
03. Digest and centrifuge the cells (refer to the subculture protocol for specific steps).
04. Resuspend the cells in cryopreservation solution at 1 mL per tube, mix thoroughly by pipetting, and aliquot into cryovials.
05. Place the cells in a programmed cooling box and freeze them in a -80°C freezer.
06. The cells are subsequently transferred to a liquid nitrogen tank for long-term storage.
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