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How can I confirm the PTMScan® LysC Protease #84748 digested my proteins efficiently?

To confirm digestion, we recommend taking a ~1% aliquot of the protein sample pre- and post-digestion and running them on an SDS-PAGE gel with Coomassie blue staining. After di...

Which Cas9 antibody is the most sensitive by western immunoblot?

The most sensitive antibody will depend on whether you are trying to detect Cas9 protein from Streptococcus pyogenes, Staphylococcus aureus, or Campylobacter jejun...

Is PTMScan® compatible with SILAC?

PTMScan® is compatible with the use of stable isotope labeling using amino acids in cell culture, or SILAC. The incorporation of heavy stable iso...

What is the concentration of the pAG-MNase Enzyme #57813?

The concentration of the pAG-MNase Enzyme #57813 in our CUT&RUN kits is 25ug/ml.

Which caspase-3 antibody do you suggest for paraffin-embedded immunohistochemistry (IHC-P) with human samples?

We have two antibodies for detecting caspase-3 that are validated for IHC-P with human samples, Caspase-3 Antibody #9662 and Caspase-3 (D3R6Y) Rabbit mAb (IHC Formulated) #14214. W...

How are the SignalStar™ Multiplex IHC Kits & Reagents validated?

CST thoroughly validates each antibody available in the

What is the expected size distribution of CUT&RUN DNA?

When using antibodies for CUT&RUN against histone modifications, we typically see fragment sizes as small as 150 bp (size of a mono-nucleosome) and quite often see a nucleosoma...

Can I reuse PTMScan® antibody beads?

The “classic” PTMScan® kits that use agarose beads and the PTMScan HS kits that use magnetic beads are both designed for a single PTM peptide enrichment. The nature of the acidic e...

Can the Cell Fractionation Kit #9038 be used for frozen cell pellets?

We have had customers perform this assay using frozen cell pellets, prior to adding any fractionation buffer, and the subsequent fractionation worked fine. Once you start fracti...

In terms of cytokine activity, what is the difference between International Units (IU) and specific activity?

International Units (IU) and specific activity are two differe...

What DNA library prep kit do you recommend using for CUT&RUN?

We recommend using our SimpleChIP® ChIP-seq DNA Library Prep Kit for Illum...

What advice do you have for collecting samples of adherent cell lines versus suspension cell lines for CUT&Tag experiments?

For CUT&Tag assays using adherent cell lines, the first step is to detach the cells from the dish. We recommend using trypsin to detach the cells. Alternatively, Accutase Cell ...

Which phospho-Akt (Ser473) antibody do you suggest for Immunofluorescence (Immunocytochemistry) with mouse samples?

We have several antibodies for detecting Akt phosphorylation at Ser473 that are validated for IF-IC with mouse tissues. These are:

  • Phospho-Akt (Ser473) (D9E) XP® ...

    Can CUT&Tag-enriched DNA be analyzed by qPCR, or do I have to do NGS to analyze my data?

    If qPCR analysis is desired, we recommend performing CUT&RUN, as CUT&Tag DNA is not compatible with qPCR. The sample incubation step at 58℃ required for tagmented DNA so...

    Does CUT&RUN show a bias toward euchromatin or heterochromatin?

    We do not observe any bias toward euchromatin or heterochromatin in our assays. In CUT&RUN, the target-specific antibody recruits the pAG-MNase and directs digestion to chromat...

    Which p44/42 MAPK (Erk1/2) antibody do you suggest for immunofluorescent immunocytochemistry (IF-IC) in human samples?

    We have several monoclonal antibodies for p44 and p42 mitogen-activated protein kinases (MAPKs), also referred to as Erk2 and Erk1, that are validated for IF-IC with human samples....

    Which phospho-Akt (Ser473) antibody do you suggest for paraffin-embedded immunohistochemistry (IHC-P) with human samples?

    We have several antibodies for detecting Akt phosphorylation at Ser473 that are validated for IHC-P with human samples. These are:

    • Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb...

      Which phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) antibody do you suggest for paraffin-embedded immunohistochemistry (IHC-P) with human samples?

      We have several antibodies for detecting p44 and p42 MAPK when phosphorylated at Thr202 and Tyr204 of Erk1, or Thr185 and Tyr187 of Erk2, that are validated for IHC-P with human sa...

      What is the intended use of PTMScan® LysC Protease #84748?

      The intended use of the PTMScan® LysC Protease #84748 is to digest proteins into peptides as input for PTMScan Kits where an arginine is part of the PTM motif sequence.

      A li...

      Do you have a methylene blue dot blot staining protocol?

      We have found that the methylene blue staining of the amounts of DNA spotted when performing a DNA dot blot can be tricky and may need optimization.

      1. It is very important ...

      Can I store the unopened, lyophilized PTMScan® LysC Protease #84748 at -80°C instead of -20°C?

      Yes, the lyophilized PTMScan® LysC Protease #84748 may be stored at -80°C. However, we do not recommend storing the LysC powder in a frost-free freezer or a freezer that undergoes ...

      Is there a maximum number of cells that can be used in a single CUT&Tag assay?

      We have found 100,000 cells per assay to be sufficient for enriching target loci for multiple histone modifications, transcription factors, and transcription cofactors. However, on...

      Do you have a protocol for lysing in a 96-well plate (no sonication)?

      For lysis in 96-well assay plates, we recommend incubation in the provided lysis buffer for ~30 min at 4°C with gentle shaking. While sonication is difficult in small volumes, w...

      What is the expected yield of a CUT&Tag DNA library?

      The expected yield of an amplified CUT&Tag DNA library depends on the DNA quantification system used.

      • If you are using a NanoDrop Spectrophotometer or QIAxpert System, ...

        Can I use the Cell Fractionation Kit #9038 with tissue samples?

        Since the Cell Fractionation Kit #9038 is primarily meant to be used for cultured cell lines, the buffers have not been optimized for use with tissue samples. Therefore, the fracti...

        What types of Ras are recognized by your G12D and G12V mutant specific antibodies?

        Our Ras (G12D Mutant Specific) (D8H7) Rabbit mAb #14429 and Ras (G12V Mutant Specific) (D2H12) Rabbit mAb #14412 will detect all three types of Ras (H-Ras, K-Ras, and N-Ras) if the...

        Will your Anti-biotin, HRP-linked Antibody #7075 cross-react with endogenous biotin in cell lysates?

        Our Anti-biotin, HRP-linked Antibody #7075 is intended for use with our Biotinylated Protein Ladder Detection Pack #7727. However, it is possible for this antibody to also detec...

        How were the sequences of the PTMScan® Control peptides chosen?

        The sequences of the synthetic peptides were based on modified peptides that we have consistently recovered and identified in human and mouse sam...

        Do your GFP antibodies detect citrine?

        We have not tested any of our GFP antibodies against the citrine variant of YFP.

        Are any of your SARS-CoV-2 Spike Proteins suitable for cell culture?

        Unfortunately, we have not tested our SARS-CoV-2 Spike Proteins in cell culture. Additionally, these products are not sterile, therefore we would not recommend using them in thi...
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