Technical Support Articles
Results (713)
Why do I see high background by western blot when probing for phospho-AMPK (Thr172)?
If CST recommended buffers are being used for western blotting and high background or additional bands persist when probing for phospho-AMPK (Thr172), the issue could be related to...
Can you provide data that confirms the specificity of your Raptor (24C12) Rabbit mAb #2280?
The specificity of our Raptor (24C12) Rabbit mAb #2280 has been confirmed by siRNA in the following papers:
PMID: 28648777 (https://pubmed.ncbi.nlm.nih.gov/28648777/) Figure...
Why do I need to use the three buffers in the Cell Fractionation Kit #9038?
Each buffer in the Cell Fractionation Kit #9038 contains detergents to help isolate different cellular fractions. The Cytoplasmic Isolation Buffer (CIB) contains a non-ionic deterg...
How should I resuspend and aliquot the PTMScan® LysC Protease #84748?
To resuspend the PTMScan® LysC Protease #84748, we recommend allowing the vial to equilibrate at room temperature for a few minutes after removing from the -20C freezer. Next, ...
How stable is the Senescence β-Galactosidase Staining Kit #9860 after multiple freeze/thaws?
We are confident that the Senescence β-Galactosidase Staining Kit #9860 will continue to perform as expected up to at least 3 freeze/thaw cycles. We recommend aliquoting your reage...
How are the SignalStar™ Multiplex IHC Kits & Reagents validated?
CST thoroughly validates each antibody available in the CST antibodies undergo thorough validation processes to ensure reliable performance for every approved application. This includes testing different protocol variations to identify ... Yes, please select “Human” in the first step of the SignalStar Multiplex IH... The SignalStar Multiplex IHC kits and reagents have been optimized with respect to fluorophore pairing and order of antibodies. As tissues vary in quality and expression level of b... 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&a... The VersicanG1-Fc (Rabbit IgG1) #15933 contained in the Hyaluronan Complete Tissue Staining Kit (HRP) #38822 and Hyaluronan Complete Tissue Staining Kit (Alexa Fluor® 488) #53721 h... We have several antibodies for detecting caspase-3 when cleaved at Asp175 that are validated for IHC-P with mouse samples. These are:
Can I modify the CST-recommended protocol for a given application?
Do you have human-reactive antibodies available for SignalStar™ Multiplex IHC?
Do I need to optimize the SignalStar™ Multiplex IHC Kits & Reagents for the type of tissue I’m using?
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??
Is the VersicanG1-Fc (Rabbit IgG1) #15933 product homologous with all species?
Which cleaved caspase-3 (Asp175) antibody do you suggest for paraffin-embedded immunohistochemistry (IHC-P) with mouse samples?
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...
Which β-actin antibody do you suggest for immunofluorescence-immunocytochemistry (IF-IC) in human samples?
We have several antibodies for detecting β-actin that are validated for IF-IC with human samples. These include:
- β-Actin (13E5) Rabbit mAb #4970
- β-Actin...
Which phospho-AMPKα antibody do you suggest for my western blot experiment?
The selection of a phospho-AMPKα antibody depends on the subunit of interest. Cell Signaling Technology has four antibodies that target phospho-AMPKα:
- Phospho-AMP...
Which cleaved caspase-3 (Asp175) antibody do you suggest for paraffin-embedded immunohistochemistry (IHC-P) with human samples?
We have several antibodies for detecting caspase-3 when cleaved at Asp175 that are validated for IHC-P with human samples. These are:
- Cleaved Caspase-3 (Asp175) Antibody #9...
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...
How does the specificity of the F(ab’)2 fragments differ from the corresponding full-length monoclonal antibodies?
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 ...
Does this antibody work for my application (western blot, immunoprecipitation, immunofluorescence, immunohistochemistry, ELISA, flow cytometry, chromatin immunoprecipitation, etc.)?
CST routinely tests antibodies in relevant applications. Those that pass rigorous application-specific validation standards are recommended. We only recommend applications that hav...
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...
Why is the concentration of my antibody so low?
At CST, our antibodies are formulated on the basis of their reactivity and performance for each approved application at a recommended dilution. For antibodies with high affinit...
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...
Does your CUT&RUN Assay Kit work with isolated nuclei?
Our CUT&RUN Assay Kit is optimized for use with whole cells and works with a large number of cell lines, both adherent and suspension. While it is not typically necessary to pr...
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
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...
What is the solution formulation of the PTMScan Control peptides?
The solution formulation of the PTMScan Control peptides is 25% acetonitrile, 74.9% water, 0.1% trifluoroacetic acid (V:V).<...
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...
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-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...