HeLa-CCC-TagRFP

Description

HeLa-Cell Cycle Chromobody®-TagRFP
HeLa cell line stably expressing Cell Cycle Chromobody® fused to the red fluorescent protein TagRFP

Specificity

VHH antibody fragment against cell cycle marker PCNA (proliferating cell nuclear antigen)

Live cell imaging of HeLa-Cell Cycle Chromobody®-TagRFP

For live cell imaging cells were seeded in ibidi µ-Slides. Time series was acquired on a spinning disk microscope (UltraVIEW VoX, PerkinElmer) equipped with a 63x/1.4 NA Plan-Apochromat oil immersion objective and a humidified, temperature- and CO2-controlled live cell chamber. TagRFP was excited with a 561 nm laser-line for typically between 100-500 ms/ image with laser intensity set to max. 10%. Confocal image z-stacks of living cells were recorded with a frame size of 1000x1000 pixels, a pixel size of 100 nm and a z-step size of 400 nm every 30 minutes. Images shown are maximum intensity projections of few mid-z-sections.

Ordering
information
SizeCodePrice
5 x 106 frozen cells in FCS free cryopreservation reagent CryoMaxxh-ccc-rplease inquire
Application
notes
  • Cell Cycle Chromobody for Real-Time Imaging and Quantification of DNA Replication in Mammalian Cells (PDF)
Related
information
Related
products

Monoclonal antibodies

Use this antibody for Western Blot and ELISA, or in IF.

Origin information

Source: human 

Tissue: cervix 

Disease: adenocarcinoma 

Cell type: epithelial 

Growth properties: adherent

Biosafety Level 1 (according to the Central Committee of Biological Safety, Germany, ZKBS)

 

Cell culture medium

Dulbecco's Modified Eagle's Medium with 4.5 g/L glucose,

110 mg/L sodium pyruvate and L-glutamine 

10% FCS (fetal bovine serum) 

50 µg/ml Gentamycin 

(optional: 1 mg/ml G418)

 

Assay medium

For live cell assays we recommend using DMEM without phenol red 

(no auto-fluorescence of phenol red ‡’ improved signal/ noise ration)

 

Thawing instructions

Cryopreserved cells can be thawed by the following procedures:

Centrifugation

  • Remove cells from storage and thaw quickly in a 37 °C water bath. We recommend eye protection by using approved safety goggles. We also suggest the use of safety gloves to protect uncovered skin.
  • Place 1 to 2 ml of thawed cells in ~25 ml of complete growth medium. Mix cell suspension gently.
  • Centrifuge the cells at ~80 x g for 2 to 3 min.
  • Check clarity of the supernatant and visibility of a consolidated cell pellet. Discard supernatant without disturbing the cells.
  • Gently resuspend the cells in complete growth medium and count the viable cells.
  • Plate the cells.

 

Direct plating

  • Remove cells from storage and thaw quickly in a 37 °C water bath. We recommend eye protection by using approved safety goggles. We also suggest the use of safety gloves to protect uncovered skin.
  • Plate cells directly with complete growth medium. Use 10 to 20 ml of complete medium per 1 ml of frozen cells.
  • Culture cells for 2 to 4 h. Replace medium with fresh complete growth medium to remove cryopreservative.

 We recommend thawing procedure 1.

 

Quality control

All cell lines supplied by ChromoTek undergo comprehensive quality control. 

This cell line tested negative for Mycoplasma using PAA MycoTrace Mycoplasma Detection Kit.

Cell viability: > 95% after thawing

Stability of transgene: > 95% after 3 months without G418 selection

Storage instructions

Shipped on dry ice. Store in liquid nitrogen. Short term storage at -80°C.

Only for research applications, not for diagnostic or therapeutic use.