Technical Support Articles
Results (689)
Is the VersicanG1-Fc (Rabbit IgG1) #15933 product homologous with all species?
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 has been tested succe...
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 affinity, optimal performan...
What controls should I include when performing a Simple Western™ experiment?
In addition to a known positive control sample, we recommend the following negative controls for your Simple Western™ experiment.
1. A no lysate control. This includes the primary antibody at t...
Is there a maximum number of cells that can be used in a single CUT&RUN assay?
We have found 100,000 cells per assay to be sufficient for enriching target loci for multiple histone modifications, transcription factors, and transcription cofactors. However, one can increase the n...
Why do I see a doublet when using my total Histone H3 antibody for western blot?
We see this doublet phenomenon often in a number of different cell lines that we test with our Histone H3 antibodies. It does appear to be sample dependent. The doublet may be due to a proteolytic fra...
Do your phospho-histone H2A.X (Ser139) antibodies detect gammaH2A.X or yH2A.X?
The Ser139 phosphorylation site in H2A.X has been widely referenced in the literature by researchers as "gammaH2A.X" or "yH2A.X". These names refer to the same protein and are inte...
Can the SARS-CoV-2 Spike Protein Serological IgG ELISA Kit #20154 be used for quantitation?
The SARS-CoV-2 Spike Protein Serological IgG ELISA Kit #20154 kit is meant to be used to determine if the sample is positive or negative for SARS-CoV-2 and not for relative quantitation. The included ...
Would the Human ACE2 (18-652) Recombinant Protein (mFc-Tag) #83986 or the Human ACE2 (multimeric) (18-652) Recombinant Protein #85054 work better in a competition assay?
Multimers are likely to demonstrate higher apparent affinity due to avidity effects. However, an exact recommendation for a competition assay would depend on other assay parameters, specifically as...
Can anti-rabbit or anti-mouse IgG HRP- or AP-linked secondary antibodies be used for IHC?
We have not attempted staining using Anti-rabbit IgG HRP-linked Antibody #7074, Anti-mouse IgG HRP-linked Antibody #7076, Anti-rabbit IgG AP-linked Antibody #7054 or Anti-mouse IgG AP-linked Antibody ...
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...
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 detergent used as a cell p...
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 reagents into single use ...
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?
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...
Do you have human-reactive antibodies available for SignalStar™ Multiplex IHC?
Yes, please select “Human” in the first step of the SignalStar Multiplex IHC Panel Builder ...
How do you make the 4X RNA Denaturing Buffer for RNA dot blot?
4X RNA Denaturing Buffer Recipe for use with our RNA dot blot protocol:
657ul Formamide @99.5%
210ul of 37% Formaldehyde
133ul of 10x MOPS buffer
Final formulation (considering rou...
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...
Which Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) antibody do you suggest for paraffin-embedded immunohistochemistry (IHC-P) with mouse 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,...
Do I need to optimize the SignalStar™ Multiplex IHC Kits & Reagents for the type of tissue I’m using?
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 biomarkers, increasin...
Should I be concerned that my Concanavalin A beads "clump together" in some of my CUT&RUN experiments?
The Concanavalin A beads can clump together rather easily, especially when the cells become lysed. To avoid this, make sure your cells are healthy and be sure to treat them very gently during the cell...
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. Our We have several antibodies for detecting β-actin that are validated for IF-IC with human samples. These include: Which β-actin antibody do you suggest for immunofluorescence-immunocytochemistry (IF-IC) in 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 solubilization in the ...
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 elution step, which l...
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.
How does the specificity of the F(ab’)2 fragments differ from the corresponding full-length monoclonal antibodies?
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 with human samples. Th...