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  7. Invitrogen™ TaqMan™ Cells-to-CT™ HepatoExpress™ Kit

Invitrogen™ TaqMan™ Cells-to-CT™ HepatoExpress™ Kit
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Catalog No. a65791
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A65791 2500 Reactions (Lysis only)
A65788 40 Reactions
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Catalog No. A65791 Supplier Invitrogen™ Supplier No. A65791

Please call Customer Service at 1-800-234-7437 or send an email to help@thermofisher.com for assistance.

Description

The TaqMan Cells-to-CT HepatoExpress Kit is a complete cell lysate system designed for gene expression analysis directly from hepatocytes without the need of RNA purification.

The TaqMan™ Cells-to-CT™ HepatoExpress Kit is a hepatocyte gene expression kit designed for RNA extraction-free RT-qPCR. It enables fast, direct lysis of primary hepatocytes, eliminating the need for RNA purification while reducing sample preparation time to ~10 minutes.

This streamlined cell-to-CT workflow allows researchers to perform high-sensitivity gene expression analysis directly from cells, without columns, beads, or centrifugation. The result is reliable, reproducible RT-qPCR data with significantly reduced hands-on time.

Optimized specifically for primary hepatocyte models—including ECM-overlay cultures—the kit supports high-throughput, automation-compatible workflows for drug metabolism, toxicity studies, and mechanistic research.

Key Benefits

Fast, Extraction-Free Gene Expression Workflow

  • Perform RT-qPCR without RNA isolation or purification
  • Reduce RNA preparation time from hours to ~10 minutes
  • Direct lysis workflow minimizes sample handling and variability

High-Quality RT-qPCR Performance

  • Sensitivity and reproducibility comparable to purified RNA workflows
  • Preserves RNA integrity during lysis
  • Compatible with multiplex qPCR and low-input samples

Optimized for Primary Hepatocytes

  • Efficient lysis of hepatocytes, including ECM-overlay cultures
  • Consistent performance across hepatocyte species and models
  • Designed for challenging in vitro liver systems

Automation-Ready and Scalable

  • Compatible with 96-well plate formats and robotic liquid handling systems
  • An ideal choice for high-throughput screening and large-scale studies
  • Room-temperature workflow simplifies lab integration

Sustainable and Simplified Workflow

  • Significantly reduces plastic consumables associated with RNA purification
  • REACH-compliant reagents support environmentally responsible lab practices

Designed for Hepatocyte Gene Expression Challenges

Primary hepatocytes are widely used for drug metabolism, CYP450 induction, and toxicity studies, but efficient RNA extraction from these cells—especially in ECM-overlay cultures—can be difficult and time-consuming.

The HepatoExpress Kit addresses these challenges with a direct lysis chemistry that:

  • Effectively penetrates extracellular matrix (ECM) overlays
  • Protects RNA during lysis without purification
  • Maintains compatibility with reverse transcription and qPCR

This enables accurate gene expression analysis without the variability introduced by traditional RNA extraction workflows.

Applications

The HepatoExpress Kit is an ideal choice for:

  • Hepatocyte gene expression analysis
  • CYP450 induction assays
  • Drug metabolism and ADME studies
  • Toxicology and safety screening
  • Target verification and early-stage compound screening
  • Mechanistic studies in liver biology

Scalable from Discovery to High-Throughput Screening

Whether performing small-scale mechanistic studies or large-scale screening, the kit supports flexible experimental design. Its automation-friendly format enables seamless integration into high-throughput workflows, improving reproducibility and throughput.

Workflow: From Cells to qPCR in ~75 Minutes

The simple three-step workflow enables rapid gene expression analysis:

  1. Cell lysis (~10 minutes, room temperature)
    • Simultaneous cell lysis, RNA preservation, and genomic DNA removal
  2. Reverse transcription
    • cDNA synthesis using SuperScript™ IV VILO™ Master Mix
  3. qPCR detection
    • Sensitive, multiplex-capable amplification using TaqMan™ Gene Expression Assays

Compatible with 700,000+ TaqMan™ Gene Expression Assays, enabling broad pathway and target coverage.

Why Choose an RNA Extraction-Free qPCR Workflow?

Compared to traditional column-based RNA isolation methods, the HepatoExpress Kit:

  • Reduces workflow time from hours to minutes
  • Minimizes sample loss and variability
  • Reduces multiple handling steps that can degrade RNA
  • Improves consistency across replicates and experiments

The TaqMan™ Cells-to-CT™ HepatoExpress Kit is a fast, reliable, and scalable solution for primary hepatocyte gene expression analysis. By enabling direct, RNA extraction-free RT-qPCR, it simplifies workflows while maintaining high data quality—making it excellent for modern drug discovery and in vitro liver research.

Specifications

Content And Storage HepatoExpress Lysis Solution (Room Temperature), HepatoExpress LysisGuard (-20°C), Express ezDNase (-20°C). For long-term storage, store HepatoExpress Lysis Buffer at 4°C.
Detection Method TaqMan
Form Liquid
Format Kit
GC-Rich PCR Performance High
PCR Method 2-step RT-qPCR
Reaction Speed 10 min.
High-throughput Compatibility Yes
Label or Dye ROX
Product Type Direct Cell Lysis to RT-qPCR Kit
Quantity 2500 Reactions (Lysis only)
Shipping Condition Frozen, Dry Ice
Target cDNA
For Use With (Application) Gene Expression
Isolation Technology Direct Cell Lysis
No. of Reactions 2500 Reactions (Lysis only)
Purification Target RNA
Sample Type Hepatocytes, HepaRG, HepG2
Starting Material Up to 100,000 Cultured Cells
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Frequently Asked Questions (FAQs)

Is the TaqMan Cells-to-CT HepatoExpress Kit compatible with 3D cultures?

The TaqMan Cells-to-CT HepatoExpress Kit has not been tested with 3D cultures.

Is the TaqMan Cells-to-CT HepatoExpress Kit compatible with 384-well tissue culture plates?

The TaqMan Cells-to-CT HepatoExpress Kit has not been tested with tissue cells in 384-well plates.

Does the ezDNase included in the TaqMan Cells-to-CT HepatoExpress Kit need to be inactivated prior to RT-qPCR?

No, ezDNase contained in the TaqMan Cells-to-CT HepatoExpress Kit is a double-strand specific DNase and will not degrade cDNA produced in the RT reaction. It is also heat-labile and will be inactivated during the heat kill step of the RT reaction.

Should I wash my cells using warm or cold wash buffer when using the TaqMan Cells-to-CT HepatoExpress Kit?

With the TaqMan Cells-to-CT HepatoExpress Kit, cells can be washed using warm or cold wash buffer according to user preference. In our evaluation, we found that using warm or chilled wash buffer yielded equivalent results.

When using the TaqMan Cells-to-CT HepatoExpress Kit, can I go directly from cells to RT-qPCR without having to purify RNA?

Yes, the TaqMan Cells-to-CT HepatoExpress Kit allows for direct RT-qPCR amplification from cell lysates without purification of RNA.

Can I use the TaqMan Cells-to-CT HepatoExpress Kit with larger microplate well sizes?

The TaqMan Cells-to-CT HepatoExpress Kit has been tested using the 96-well plate format using the standard 50 μL lysis reaction. If larger well sizes are desired, we recommend scaling up the volume of lysis reaction accordingly and performing a validation experiment.

Can the TaqMan Cells-to-CT HepatoExpress Kit protocol be automated?

Yes, the lysate preparation step in the TaqMan Cells-to-CT HepatoExpress Kit protocol is completed at room temperature and does not require any centrifugation steps, making it very automation friendly. We recommend including an orbital shaker on your liquid handler deck for optimal performance.

Does the TaqMan Cells-to-CT HepatoExpress Kit contain reagents for the removal of genomic DNA (gDNA)?

Yes, the TaqMan Cells-to-CT HepatoExpress Kit includes Express ezDNase which is added to the lysis mixture for removal of gDNA during cell lysis. Express ezDNase is a double-strand specific, heatlabile DNase that will only digest gDNA, making it compatible with the downstream reverse transcription reaction. The enzyme is automatically inactivated during a heat kill step included in the reverse transcription program. To avoid the detection of gDNA, we recommend using a TaqMan Gene Expression Assay specifically designed to span an intron-exon boundary. Such assays are designated with an "_m1" suffix.

How do I identify genomic DNA (gDNA) contamination when using the TaqMan Cells-to-CT HepatoExpress Kit?

gDNA contamination can be identified by performing an RT control reaction without the RT enzyme. The TaqMan Cells-to-CT HepatoExpress Kit contains the SuperScript IV VILO Master Mix No-RT Control which includes all components except for the RT enzyme and can be used for monitoring of gDNA contamination. To avoid the detection of gDNA, we recommend using a TaqMan Gene Expression Assay specifically designed to span an intron-exon boundary. Such assays are designated with an "_m1" suffix.

Can I freeze and store cell lysates from the TaqMan Cells-to-CT HepatoExpress Kit?

Yes, lysates from theTaqMan Cells-to-CT HepatoExpress Kit are stable at -80°C for at least 1 year. We have also shown that lysates can go through five freeze/thaw cycles without any loss in signal. It is important to thaw frozen lysates on ice and keep on ice once thawed.

Can I freeze and store cDNA generated with the TaqMan Cells-to-CT HepatoExpress Kit?

Yes, cDNA generated with the TaqMan Cells-to-CT HepatoExpress Kit is stable at -80°C for at least 1 year. We have also shown that cDNA can go through five freeze/thaw cycles without any loss in signal.

Are the components of the TaqMan Cells-to-CT HepatoExpress Kit sold separately?

Yes. The lysis reagents within the TaqMan Cells-to- CT HepatoExpress Kit can be purchased in bulk under Cat. No. A65791. The standalone SuperScript IV VILO Master Mix (Cat. No. 11756050), and TaqMan Fast Advanced Master Mix for qPCR (Cat. No. 4444556) can also be purchased separately.

Can the TaqMan Cells-to-CT HepatoExpress Kit be used for multiplexed qPCR?

Yes, the TaqMan Fast Advanced Master Mix for qPCR included in the TaqMan Cells-to-CT HepatoExpress Kit supports multiplexed qPCR reactions.

What is the optimal number of primary hepatocytes to use in the lysis reaction with the TaqMan Cells-to-CT HepatoExpress Kit?

The acceptable range of cell inputs to use with the TaqMan Cells-to-CT HepatoExpress Kit per well of a 96-well plate are:
- Human primary hepatocytes: up to 100,000 cells
- Mouse primary hepatocytes: up to 19,500 cells
- Rat primary hepatocytes: up to 45,000 cells
- HepaRG: up to 100,000 cells
- HepG2: up to 100,000 cells
Does the TaqMan Cells-to-CT HepatoExpress Kit protocol have any stopping points?

Yes, the TaqMan Cells-to-CT HepatoExpress Kit allows for the following stopping points:
- After cell lysis: Cell lysates are stable at room temperature for up to 30 minutes, or can be safely stored on ice for up to 2 hours. For long term storage, lysates may be frozen at -80°C for up to 1 year. It is important to thaw frozen lysates on ice and keep on ice once thawed.
- After assembly of the RT reaction: Assembled RT reactions can be stored on ice for up to 9 hours
- After completion of the RT reaction: Completed RT reactions can be stored at -80°C for up to 1 year
- After assembly of the qPCR reaction: Assembled qPCR reactions can be stored at room temperature for up to 72 hours

What are the advantages of the two-step RT-qPCR workflow of the TaqMan Cells-to-CT HepatoExpress Kit?

The two-step RT-qPCR workflow of the TaqMan Cells-to-CT HepatoExpress Kit enables the user to reverse transcribe the entire transcriptome in one RT reaction and then perform separate qPCR reactions for any gene of interest. Two-step RT-qPCR provides the ability to archive cDNA, analyze multiple genes, and gives greater flexibility.

What is the qPCR master mix provided in the TaqMan Cells-to-CT HepatoExpress Kit?

The TaqMan Cells-to-CT HepatoExpress Kit includes the TaqMan Fast Advanced qPCR Master Mix (Cat. No. 4444965). The master mix can be purchased separately if larger qPCR volumes are required by the user.

Which reverse transcriptase is included in the TaqMan Cells-to-CT HepatoExpress Kit?

The TaqMan Cells-to-CT HepatoExpress Kit includes the SuperScript IV VILO Master Mix (Cat. No. 11756050). The master mix can be purchased separately if larger RT volumes are required by the user.

Which real-time PCR instruments are compatible with the TaqMan Cells-to-CT HepatoExpress Kit?

The TaqMan Cells-to-CT HepatoExpress Kit contains TaqMan Fast Advanced qPCR Master Mix and is compatible with the same instruments listed for the TaqMan Fast Advanced qPCR Master Mix, e.g., QuantStudio 3, 5, 6 Pro, 7 Pro, 6 Flex, 7 Flex and 12K Flex instruments. We have tested the TaqMan Cells-to-CT HepatoExpress Kit with the QuantStudio 5 instrument.

Can I use the TaqMan Cells-to-CT HepatoExpress Kit with TaqMan Arrays?

Yes, the TaqMan Cells-to-CT HepatoExpress Kit is compatible with TaqMan Array plates and cards. We suggest using the recommended input of cDNA as outlined in the TaqMan Array plates and cards protocols.

What primers are used for reverse transcription in the TaqMan Cells-to-CT HepatoExpress Kit?

The TaqMan Cells-to-CT HepatoExpress Kit includes the SuperScript IV VILO Master Mix which contains oligo (DT)18 and random hexamer primers. This primer mix will reverse transcribe all cellular RNAs.

Does the TaqMan Cells-to-CT HepatoExpress Kit include a control for RT reactions?

Yes, the TaqMan Cells-to-CT HepatoExpress Kit includes the SuperScript IV VILO Master Mix No-RT Control which can be used to determine if the sample contains genomic DNA. For a positive control, we recommend assaying for a highly expressed housekeeping gene.

Can I store the HepatoExpress Lysis Solution included in the TaqMan Cells-to-CT HepatoExpress Kit at room temperature?

Yes, the HepatoExpress Lysis Solution in the TaqMan Cells-to-CT HepatoExpress Kit can be stored at room temperature or 4°C.

Is the TaqMan Cells-to-CT HepatoExpress Kit eco-friendly?

Yes, every component of the TaqMan Cells-to-CT HepatoExpress Kit is REACH (Registration, Evaluation, Authorisation and Restriction of Chemicals)-compliant and non-hazardous. Additionally, there is a significant reduction in plastic waste compared to using column-purified RNA.

How does the performance of the TaqMan Cells-to-CT HepatoExpress Kit compare to purified RNA?

The TaqMan Cells-to-CT HepatoExpress Kit consistently produces results that are comparable to those obtained using column-purified RNA over a diverse set of target genes and cell inputs. Comparison data can be found on the product page.

How does the performance of the TaqMan Cells-to-CT HepatoExpress Kit compare to other Cells-to-CT kits?

The TaqMan Cells-to-CT HepatoExpress Kit helps provide higher sensitivity than other Cells-to-CT kits when used with primary hepatocytes.

What is the purpose of the LysisGuard in the TaqMan Cells-to-CT HepatoExpress Kit?

The LysisGuard contained in the TaqMan Cells-to- CT HepatoExpress Kit works in conjuction with the HepatoExpress Lysis Solution to stabilize the sample.

How does the HepatoExpress Lysis Solution contained in the TaqMan Cells-to-CT HepatoExpress Kit differ from lysis buffers from other Cells-to-CT kits?

The HepatoExpress Lysis Solution contained in the TaqMan Cells-to-CT HepatoExpress Kit has been formulated to work specifically with primary hepatocytes and with extracellular matrix (ECM) overlays. This solution does not require the use of a stop solution, and is REACH (Registration, Evaluation, Authorisation and Restriction of Chemicals)-compliant and free of hazardous materials.

What is the maximum amount of cDNA that I can use in the qPCR reaction using the TaqMan Cells-to-CT HepatoExpress Kit?

With the TaqMan Cells-to-CT HepatoExpress Kit, up to 40% of the qPCR reaction volume can be cDNA.

What is the maximum amount of cell lysate that I can use in the RT reaction with the TaqMan Cells-to-CT HepatoExpress Kit?

With the TaqMan Cells-to-CT HepatoExpress Kit, up to 10% of the RT reaction volume can be cell lysate.

What extracellular matrices (ECMs) are compatible with TaqMan Cells-to-CT HepatoExpress Kit?

We have tested Thermo Fisher Scientific’s Gibco Geltrex LDEV-Free Reduced Growth Factor Basement Membrane Matrix (Cat. No. A1413201) and Corning Matrigel Matrix (Cat. No. 356237) with the TaqMan Cells-to-CT HepatoExpress Kit. We anticipate that the kit will be broadly compatible with other similar ECM products but they should be evaluated by the user.

Can I scale down the volume of the lysis reaction with the TaqMan Cells-to-CT HepatoExpress Kit?

We have not evaluated the use of smaller lysis reaction volumes with the TaqMan Cells-to-CT HepatoExpress Kit and recommend using the standard 50 μL lysis reaction volume.

Can I use the TaqMan Cells-to-CT HepatoExpress Kit with other types of cells besides primary hepatocytes?

We have not evaluated the use of other cells with the TaqMan Cells-to-CT HepatoExpress Kit. We recommend performing a test experiment for any cell types beyond primary hepatocytes, HepaRG, and HepG2.

Should I be concerned if I see precipitates in the TaqMan Fast Advanced qPCR Master Mix?

The TaqMan Fast Advanced qPCR Master Mix can develop particulates that float in solution. Such particulates do not affect kit performance. To minimize the formation of particulates, the master mix should be initially thawed at room temperature and then stored at 4°C for long-term storage.

For Research Use Only. Not for use in diagnostic procedures.