Technical Support Articles
Results (689)
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 stability tested by ex...
Do you have mouse-reactive antibodies available for SignalStar™ Multiplex IHC?
Yes, please select “Mouse” in the first step of the SignalStar Multiplex IHC Panel Builder ...
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 control, which may som...
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 antibodies work in the CUT&am...
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 isotopes has no effect ... 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 completion of staini... SignalStar Multiplex IHC kits and reagents haven’t yet been validated for use with antibodies outside of our menu. We’re in the process of developing cust... 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 possible to perform ... 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 that the correct sub... The molar extinction coefficient of rabbit IgG at 280 nm is approximately 210,000 M-1cm-1. 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?
How long after the completion of staining can I wait to image my slides when performing a SignalStar™ Multiplex IHC assay?
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?
Can my slides be used for other assays after SignalStar™ Multiplex IHC?
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?
What is the extinction coefficient of my rabbit antibody?
Which cleaved caspase-3 (Asp175) antibody do you suggest for immunofluorescent immunocytochemistry (IF-IC) with mouse samples?
Can I modify the CST-recommended protocol for a given application?
CST antibodies undergo thorough validation processes to ensure reliable performance for every approved application. This includes testing different protocol variations to identify the most effective o...
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 because the emission spe...
What are the differences between the BrdU Cell Proliferation Chemiluminescent Assay Kit #5492 and the BrdU Cell Proliferation Assay Kit #6813? Which should I choose?
In general, the BrdU Cell Proliferation Chemiluminescent Assay Kit #5492 offers a broader dynamic range and higher sensitivity compared to the BrdU Cell Proliferation Assay Kit #6813, which is colorim...
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 protein:
- I...
What controls can I use to show my CUT&RUN experiment is working?
Whether performing CUT&RUN with downstream qPCR or NG-seq analysis, we always recommend using a CUT&RUN-validated positive control antibody like our 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:
Which phospho-Akt (Ser473) antibody do you suggest for Immunofluorescence (Immunocytochemistry) (IF-IC) with human samples?
Can I use primers from the Illumina Nextera Library preparation system or design my own primers to amplify the DNA library generated from the CUT&Tag Assay Kit #77552??
Yes, you can use primers from the Illumina Nextera Library preparation system or self-designed primers to amplify the DNA library generated from the CUT&Tag DNA using the CUT&Tag Assay Kit #77...
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 product webpage or to...
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. We recommend #14214 f...
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 digestion, you should ...
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 not show cross-reac...
Can I store the resuspended PTMScan® LysC Protease #84748 aliquots at -20°C instead of -80°C?
Although the lyophilized powder can be stored at -20°C, we recommend long-term storage of the resuspended LysC at -80°C. We do not recommend long-term storage of the resuspended LysC at -20°C due to p...
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 chromogenic IHC, denoted ...
Why do my Concanavalin A Magnetic Beads #82307 appear aggregated?
As long as your Concanavalin A Magnetic Beads #82307 have been stored at the appropriate recommended temperature, the appearance of aggregation is not a cause for concern. By nature, the bead particle...
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 5,000-10,000 cells for histo...
Is it possible to perform CUT&Tag on samples like heart, brain, or developing mouse tissues?
Yes. We have shown that our CUT&Tag Assay Kit #77552 works with mouse liver, mouse brain, and mouse heart tissues for histone targets with as little as 1 mg of tissue for each CUT&Tag reaction...
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 #4685
- Akt Antibo...
Which cleaved caspase-3 (Asp175) antibody do you suggest for immunofluorescent immunocytochemistry (IF-IC) with human samples?
We have several antibodies for detecting Cleaved Caspase-3 (Asp175) that are validated for IF-IC with human samples. These are:
- Cleaved Caspase-3 (Asp175) Antibody #9661
- Cleaved Caspas...