Technical Support Articles

Do you have mouse-reactive antibodies available for SignalStar™ Multiplex IHC?

What is the fewest number of cells I can use in a reaction with the CST CUT&Tag kit?

We have shown that our CUT&Tag Assay Kit #77552 works with as few as 5,000-10,000 cells for histone modifications and 20,000 cells for transcription factors and cofactors.

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

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

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

Can CUT&RUN enriched DNA be analyzed by qPCR, or do I have to do NGseq to analyze my data?

What is the best way to choose an antibody for my CUT&RUN assay if I cannot find a CUT&RUN validated antibody for my target protein?

When selecting an antibody for use in CUT&RUN, we recommend starting with a ChIP or ChIP-seq-validated antibody; however, we have found that not all ChIP-validated antibodie...

What is an appropriate positive control to include in this SignalStar™ assay? Are multiple controls necessary?

Any tissue shown to be positive for each marker via chromogenic IHC can serve as a positive control tissue for a SignalStar™ assay. Each target will therefore require a positive co...

Which nuclear dyes can be used for In-Cell Western assays in combination with DyLight 680-conjugated secondary antibodies?

When performing In-Cell Western assays with DyLight 680-conjugated secondary antibodies, we recommend using either Hoechst 33342 or DAPI as a nuclear dye. DRAQ5 cannot be used beca...

Have species been tested if they are not listed as cross-reactive with a CST antibody on the datasheet?

Possibly. CST scientists test all our antibodies on multiple species when feasible, depending on the target protein and the experimental systems available. If a tested species does...

What is the extinction coefficient of my rabbit antibody?

The molar extinction coefficient of rabbit IgG at 280 nm is approximately 210,000 M-1cm-1.

What are the two bands seen in western blot analysis using ANKRD15 (E9I4I) Rabbit mAb #69953?

When using ANKRD15 (E9I4I) Rabbit mAb #69953, two distinct bands are typically observed in western blotting experiments. These bands represent two different isoforms of the ANKRD15...

How should I store my antibody?

Proper storage of your antibody may vary depending on the product's storage buffer and antibody conjugation. Therefore, please refer to the "Storage" section of the p...

Do I need to perform Fc blocking when using rabbit or mouse antibodies in flow cytometry?

Human Fc-gamma Receptor 1a (FCGR1A) exhibits strong cross-reactivity with rabbit IgG, and mouse IgG2a, IgG2c, and IgG3. Of course, mouse Fc-gamma Receptors will react with mouse an...

Which method is better for purifying CUT&Tag DNA, DNA affinity columns or a phenol-chloroform DNA extraction using ethanol precipitation?

We recommend using DNA Purification Buffers and Spin Columns (ChIP, CUT&RUN, CUT&Tag) #14209 when purifying your CUT&Tag DNA. Although phenol-chloroform extraction foll...

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...

Why does the material in my new vial of Echinomycin #51434 look different?

Echinomycin #51434 is lyophilized into the vial, and any difference in the appearance of the lyophilized echinomycin is due to a dry coating of the compound in the vial. When you a...

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...

Which phospho-S6 ribosomal protein (Ser235/236) antibody do you suggest for immunofluorescent immunocytochemistry (IF-IC) 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,<...

Which Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) antibody do you suggest for immunofluorescent immunocytochemistry (IF-IC) with human samples?

We have several monoclonal 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 IF-IC wi...

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....

My items say to store at -20°c but they were shipped in an envelope. Is it ok to use?

Yes, it is safe to use these items. Since 2003, CST has been shipping antibodies at room temperature when possible to reduce waste from packing materials. All CST antibodies are st...

I don’t see my target of interest in your menu of available antibodies for SignalStar™ Multiplex IHC. Can I still use it in my panel in some way?

SignalStar Multiplex IHC kits and reagents haven’t yet been validated for use with antibodies outside of our menu. We’re in the proces...

Can I combine antibodies used in a SignalStar™ assay with direct conjugates?

SignalStar™ Multiplex IHC Kits & Reagents have not been validated for use in combination with other assays. The SignalStar technology does not destroy the tissue, so it may be ...

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

We have several antibodies for detecting the phosphorylation of Akt at Ser 473 that are validated for IF-IC with human samples. These include the following: