Technical Support Articles
Results (709)
Do you have mouse-reactive antibodies available for SignalStar™ Multiplex IHC?
Yes, please select “Mouse” in the first step of the SignalStar Multiplex IH...
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 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... 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 ... During the course of optimization, we’ve found that fluorescent staining may show higher %-positivity than chromogenic staining. To ensure any excess staining is specific, confirm ... PTMScan® is compatible with the use of stable isotope labeling using amino acids in cell culture, or SILAC. The incorporation of heavy stable iso... 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... 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 ... We have several antibodies for detecting caspase-3 when cleaved at Asp175 that are validated for IF-IC with mouse samples. These are:
My items say to store at -20°c but they were shipped in an envelope. Is it ok to use?
How long after the completion of staining can I wait to image my slides when performing a SignalStar™ Multiplex IHC assay?
When comparing my SignalStar™ staining to the chromogenic staining on a serial section, I see more positive cells. How do I know if this excess staining is correct?
Is PTMScan® compatible with SILAC?
What is an appropriate positive control to include in this SignalStar™ assay? Are multiple controls necessary?
Can my slides be used for other assays after SignalStar™ Multiplex IHC?
Which cleaved caspase-3 (Asp175) antibody do you suggest for immunofluorescent immunocytochemistry (IF-IC) with mouse samples?
Can I autoclave the 6-Tube Magnetic Separation Rack #7017?
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 ...
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...
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 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...
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...
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.
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...
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...
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...
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...
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...
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...
Which total Akt antibody do you suggest for paraffin-embedded immunohistochemistry (IHC-P) with mouse samples?
We have several monoclonal antibodies for detecting total Akt protein that are validated for IHC-P with mouse samples. These are as follows:
- Akt (pan) (C67E7) Rabbit mAb #...
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...
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...
Which total Akt antibody do you suggest for paraffin-embedded immunohistochemistry (IHC-P) with human samples?
We have several monoclonal antibodies for detecting total Akt protein that are validated for IHC-P in human samples. These are as follows:
- Akt (pan) (C67E7) Rabbit mAb #46...
What conditions do you recommend for primary antibody incubation in my IF experiment?
We recommend dilution of the primary antibody into PBS containing 1% BSA and 0.3% Triton X-100. BSA acts as a carrier protein and improves antibody stability in dilute environments...
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...