KC-6720

MIAPaCa-2-PAN5-6-Low Cell Line

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Background of MIAPaCa-2-PAN5-6-Low Cell Line

Survivin aliases is BIRC5. BIRC5 (Baculoviral IAP Repeat Containing 5) is a Protein Coding gene.This gene is a member of the inhibitor of apoptosis (IAP) gene family, which encode negative regulatory proteins that prevent apoptotic cell death. IAP family members usually contain multiple baculovirus IAP repeat (BIR) domains, but this gene encodes proteins with only a single BIR domain. The encoded proteins also lack a C-terminus RING finger domain. Gene expression is high during fetal development and in most tumors, yet low in adult tissues. Alternatively spliced transcript variants encoding distinct isoforms have been found for this gene.P53 mutations are universal across cancer types.This gene encodes a tumor suppressor protein containing transcriptional activation, DNA binding, and oligomerization domains. The encoded protein responds to diverse cellular stresses to regulate expression of target genes, thereby inducing cell cycle arrest, apoptosis, senescence, DNA repair, or changes in metabolism. Mutations in this gene are associated with a variety of human cancers, including hereditary cancers such as Li-Fraumeni syndrome. Alternative splicing of this gene and the use of alternate promoters result in multiple transcript variants and isoforms. Additional isoforms have also been shown to result from the use of alternate translation initiation codons from identical transcript variants.Ras k 1 is a major allergen from the Indian mackerel (Rastrelliger kanagurta). It belongs to the parvalbumin family, a calcium-binding protein highly conserved in fish muscles. With a molecular weight of ~11.3 kDa, Ras k 1 is heat-stable and responsible for IgE-mediated allergic reactions in fish-sensitive individuals.

Specifications

Catalog NumberKC-6720
Cell Line NameMIAPaCa-2-PAN5-6-Low Cell Line
Clone Number1#
Host Cell LineMIAPaCa-2
DescriptionMIAPaCa-2 cell line stably expressing exogenous PAN5-6 gene
QuantityTwo vials of frozen cells (≥2-106/vial)
StabilityStable in culture over a minimum of 10 passages
ApplicationDrug screening and biological assays
Freezing Medium70% DMEM + 20% FBS + 10% DMSO
Propagation MediumDMEM+10%FBS+2.5%HS+1μg/mL puromycin
Selection MarkerPuromycin
MorphologyEpithelial
SubcultureSplit saturated culture 1:4-1:6 every 2-3 days; seed out at about 1-3 × 105 cells/mL
Incubation37 °C with 5% CO2
Doubling TimeApproximately 30 hours
Mycoplasma StatusNegative

Cell Line Generation

MIAPaCa-2-PAN5-6-low cell line was generated using a lentiviral vector expressing the PAN5-6 sequence.

Characterization

Figure1: Characterization of PAN5-6 overexpressing in MIAPaCa-2-PAN5-6-low stable clones using WB.

Figure2: Characterization of MIAPaCa-2-PAN5-6-low cell line stable clone using PCR sequencing.

Cell Resuscitation

  1. Prewarm culture medium (DMEM+10%FBS+2.5%HS+1μg/mL puromycin) in a 37°C water bath.
  2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
  3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
  4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
  5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
  6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
  7. Incubate the flask at 37°C, 5% CO2 incubator.
  8. Split saturated culture 1:4-1:6 every 2-3 days; seed out at about 1-3 × 105 cells/mL.

Cell Freezing

  1. Prepare the freezing medium (70% DMEM + 20% FBS + 10% DMSO) fresh immediately before use.
  2. Keep the freezing medium on ice and label cryovials.
  3. Transfer cells to a sterile, conical centrifuge tube, and count the cells.
  4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
  5. Resuspend the cells at a density of at least 3×106 cells/mL in chilled freezing medium.
  6. Aliquot 1 mL of the cell suspension into each cryovial.
  7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
  8. Transfer vials to liquid nitrogen for long-term storage.

References

  1. Petrarca CR, Brunetto AT, Duval V, Brondani A, Carvalho GP, Garicochea B. Survivin as a predictive biomarker of complete pathologic response to neoadjuvant chemotherapy in patients with stage II and stage III breast cancer. Clin Breast Cancer. 2011 Apr;11(2):129-34. doi: 10.1016/j.clbc.2011.03.002. Epub 2011 Apr 11. PMID: 21569999.
  2. Ben-Izhak O, Laster Z, Akrish S, Muska E, Gan S, Nagler RM. The salivary tip of the p53 mutagenesis iceberg: novel insights. Cancer Biomark. 2009;5(1):23-31. doi: 10.3233/CBM-2009-0567. PMID: 19242059; PMCID: PMC12922806.

Use License Agreement

Research Use Only.
Not for use in diagnostic procedures or therapeutic applications.
Redistribution of the cell line or its derivatives is prohibited without prior written permission from Kyinno Biotechnology.
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