Technical Support Articles
Results (713)
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...
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...
How long after the completion of staining can I wait to image my slides when performing a SignalStar™ Multiplex IHC assay?
For Imaging Round 1, the staining should show robust signal when imaged up to 8 hours post completion of staining. For Imaging Round 2, imaging should be performed as close to the ...
Do you have mouse-reactive antibodies available for SignalStar™ Multiplex IHC?
Yes, please select “Mouse” in the first step of the SignalStar Multiplex IH...
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...
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 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:
- Phospho-Akt (Se...
Can I combine antibodies used in a SignalStar™ assay with direct conjugates?
SignalStar Multiplex IHC kits and reagents have been validated for use in combination with direct conjugates. The SignalStar™ Fluorescence Removal Kit PTMScan® is compatible with the use of stable isotope labeling using amino acids in cell culture, or SILAC. The incorporation of heavy stable iso... No, these racks cannot be autoclaved. Exposing them to high autoclave temperatures may cause the magnets to lose their magnetism. Instead, we suggest submerging or spraying the ... 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 ... The molar extinction coefficient of rabbit IgG at 280 nm is approximately 210,000 M-1cm-1. 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... 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... We have several antibodies for detecting caspase-3 when cleaved at Asp175 that are validated for IF-IC with mouse samples. These are:
Is PTMScan® compatible with SILAC?
Can I autoclave the 6-Tube Magnetic Separation Rack #7017?
Can my slides be used for other assays after SignalStar™ Multiplex IHC?
What is the extinction coefficient of my rabbit antibody?
Which nuclear dyes can be used for In-Cell Western assays in combination with DyLight 680-conjugated secondary antibodies?
What are the two bands seen in western blot analysis using ANKRD15 (E9I4I) Rabbit mAb #69953?
Which cleaved caspase-3 (Asp175) antibody do you suggest for immunofluorescent immunocytochemistry (IF-IC) with mouse samples?
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...
What is the fewest number of cells I can use in a reaction with your CUT&RUN Assay Kit #86652?
We have shown that our CUT&RUN Assay Kit #86652 works with as few as 5,000-10...
Which total Akt antibody do you suggest for immunofluorescence-immunocytochemistry (IF-IC) with mouse samples?
We have several total Akt antibodies that are validated for IF-IC with mouse samples. These are:
- Akt (pan) (C67E7) Rabbit mAb #4691
- Akt (pan) (11E7) Rabbit mAb #468...
Can I use CST antibodies for fluorescent detection in FFPE tissue?
At CST we do not typically validate antibodies using indirect fluorescent detection in formalin fixed, paraffin embedded (FFPE) tissue. If a product is recommended for use in chrom...
What is the most suitable control DNA sample for my CUT&RUN NG-seq experiment?
When performing CUT&RUN with downstream NG-seq analysis, one can use a normal IgG antibody enriched sample as the negative control. However, we have seen and heard from other s...
Can CUT&RUN enriched DNA be analyzed by qPCR, or do I have to do NGseq to analyze my data?
Our CUT&RUN Assay Kit #86652 is compatible with downstream q...
How much CUT&RUN DNA is required for NG-seq analysis?
The amount of CUT&RUN DNA required for NG-seq analysis depends on the sensitivity of the DNA library prep kit. The typical yield for CUT&RUN DNA is 0.6 to 6 ng per reaction...
Can I use PTMScan® LysC Protease #84748 for tandem mass tag (TMT) labeling?
We have used LysC in conjunction with trypsin for TMT-labeling-based studies to reduce miscleavages. For TMT-based workflows, we recommend a higher LysC:substrate ratio of 1:100 co...
Why is the PTMScan® LysC Protease #84748 digestion performed at room temperature and not at 37°C?
The digestion is performed at room temperature to minimize carbamylation (also known as carbamoylation) of the sample peptides in the urea buffer, which can interfere with downstre...
What are the specifications for the 96-well plate in the CST ELISA kits?
For the 96-well colorimetric PathScan® and FastScan® ELISA plates, the specification information can be found in the We have several monoclonal antibodies for detecting total Akt protein that are validated for IHC-P in human samples. These are as follows:
Which total Akt antibody do you suggest for paraffin-embedded immunohistochemistry (IHC-P) with human samples?
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...
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...
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 ...
Do you recommend performing size selection of the DNA during the library preparation for CUT&Tag experiments?
No, we don't recommend performing size selection during the library purification for CUT&Tag. Based on our experience and feedback from customers, size selection can signif...