Technical Support Articles
Results (711)
Does your Anti-mouse IgG, HRP-linked Antibody #7076 cross react with mouse IgM?
Our Anti-mouse IgG, HRP-linked Antibody #7076 shows approximately 25% reactivity to mouse IgM.
Does your Ki-67 (D2H10) Rabbit mAb #9027 cross-react with mouse tissue by IHC?
We have tested the Ki-67 (D2H10) Rabbit mAb #9027 on a variety of FFPE mouse tissues and have not observed specific staining. Based on this, we have concluded that #9027 does not d...
How do the Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411 and the Phospho-Tyrosine (P-Tyr-1000) MultiMab™ Rabbit mAb mix #8954 compare in terms of their sensitivity and specificity?
The Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411 is a mouse mAb from a single clone, whereas the Phospho-Tyrosine (P-Tyr-1000) MultiMab™ Rabbit mAb mix #8954 is a mix of rabbit mAb...
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 phospho-Akt (Ser473) antibody do you suggest for Immunofluorescence (Immunocytochemistry) with mouse samples?
We have several antibodies for detecting Akt phosphorylation at Ser473 that are validated for IF-IC with mouse tissues. These are:
- Phospho-Akt (Ser473) (D9E) XP® ...
Will the SignalStain® DAB Substrate Kit #8059 still perform as expected if the material has been frozen?
While the SignalStain® DAB Substrate Kit #8059 does have a recommended storage temperature of 4°...
What is your recommended procedure for antigen retrieval with a pressure cooker?
For antigen retrieval in a pressure cooker, bring slides submerged in buffer up to 125C, maintain for 30 seconds, then cool to 90C and maintain that for 10 sec. Remove the containe...
What is the difference between phospho-Smad2 (Ser465/467) and phospho-Smad2 (Ser245/250/255)?
Phosphorylation of serines 465 and 467 indicates Smad2 activation and nuclear localization; whereas, phosphorylation of serines 245, 250, and 255 is inhibitory and leads to retenti...
Does the CST CUT&Tag kit work with isolated nuclei?
Our CUT&Tag kit is optimized for use with whole cells and works with a large number of cell lines, both adherent and suspension. While our kit works with pre-isolated nuclei be...
What is the best way to choose an antibody for my CUT&Tag assay if I can not find a CUT&Tag-validated antibody for my target protein?
We always recommend starting with CST CUT&Tag-validated antibodies because CST scientists perform intensive in-house tests to exclude any antibody that tends to show non-specif...
My Tris-Glycine SDS Running Buffer (10X) #4050 has turned yellow. Is it still okay to use?
It is normal for the Tris-Glycine SDS Running Buffer (10X) #4050 to turn yellow as it ages. The yellowing has no effect on the product's performance and is no cause for concern...
How many assays can be performed with the Senescence β-Galactosidase Staining Kit #9860?
The reagents provided with our Senescence β-Galactosidase Staining Kit #9860 are sufficient to stain 125 x 35 mm wells. The volumes listed in our protocol are intended for one ...
Will the RAG1 (D36B3) Rabbit mAb #3968 cross-react with human RAG2?
Our RAG1 (D36B3) Rabbit mAb #3968 was produced using a recombinant protein fragment with a sequence toward the N-terminal region of human RAG1. No significant homology is found in ...
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...
How do I pool CUT&Tag DNA libraries with a variety of yields?
Usually, the CUT&Tag DNA libraries from histone targets have a higher concentration than those from non-histone targets. We use the following formula to convert a library conce...
Why should I use your recommended protocol or reagents for my IHC experiment?
We have performed a number of comparisons over the years looking at protocol variables such as the retrieval solution, diluent, incubation, and detection methodology. We have seen ...
Why doesn't PD-L2 migrate at 31 kDa?
The predicted molecular weight of PD-L2 is 31 kDa (based on amino acid composition). However, in most models, PD-L2 migrates at an observed molecular weight of 45-60 kDa on Tris-Gl...
What β-Tubulin isoforms and subtypes are detected by CST β-Tubulin antibodies?
Based on homology, our β-Tubulin Antibody #2146, β-Tubulin (9F3) Rabbit mAb #2128, β-Tubulin (D3U1W) Mouse mAb #86298, and β-Tubulin (D2N5G) Rabbit mAb #15115 should detect β-Tubul...
What is the pH of SignalStain® Antibody Diluent #8112?
The pH of our SignalStain® Antibody Diluent #8112 has a range between 7.38-7.45.
Why does the observed size versus the theoretical size of TET2 differ by western blot?
TET2 is post-translationally modified (https://www.phosphosite.org/proteinAction.action?id=22594&showAllSites=true). Although the theoretical molecular weight is 223.8 kDa thes...
Which species does the Caspase-3 Activity Assay Kit #5723 work in?
We have validated the Caspase-3 Activity Assay Kit #5723 in human and mouse cell lines. However, as long as there is active caspase-3 present, this kit is predicted to work with mo...
Can you share your protocol for preparing Jurkat -/+ Anti-CD3 antibody (10 µg/mL for 2 min) cell extracts?
We prepare Jurkat -/+ Anti-CD3 antibody (10 µg/mL for 2 min) cell extracts as follows:
- Jurkat cells are cultured to the desired volume (200-400 mL) and a density o...
Can I use human PBMCs with Senescence β-Galactosidase Staining Kit #9860?
CST scientists have not tested Senescence β-Galactosidase Staining Kit #9860 using human peripheral blood mononuclear cells (PBMCs). Therefore, we cannot provide a suggested...
How can I distinguish mouse or human cells by IHC in a xenograft setting?
Our COX IV (3E11) Rabbit mAb #4850 will specifically label human cells and not mouse, and our COX IV (D6I4K) Rabbit mAb (Rodent Specific) #38563 will specifically label mouse cells...
CST provides several products to avoid detection of IgG heavy chains. Which one is best for my IP experiment?
When performing immunoprecipitation (IP) followed by western blotting, the denatured IgG light and heavy chains of the primary antibody used for IP run at approximately 25 and 50 k...
How should tissue samples be prepared for the PTMScan® Acetyl-Lysine Motif [Ac-K] Kit #13416?
Tissue samples should be placed in 1 ml of urea lysis buffer per 100 mg of tissue wet weight. For example, a 300mg tissue sample would be homogenized in 3mls of urea lysis buffer. ...
What is the minimal amount of tissue required for a CUT&RUN experiment?
While we recommend 1mg of tissue per assay in most cases, we have had some success with as little as 0.5 mg of tissue per assay. Easier tissue types, such as liver, or easier targe...
What is the concentration of the PTMScan® Ubiquitin Branch Motif (K-ε-GG) Immunoaffinity Beads #1990 component in the PTMScan® Ubiquitin Remnant Motif (K-ε-GG) Kit #5562?
The concentration of the antibody-beads included in the PTMScan® Ubiquitin Remnant Motif (K-ε-GG) Kit #5562 (Bead SKU #1990) is proprietary and therefore we are unable to disclose ...
Can your Protease/Phosphatase Inhibitor Cocktail (100X) #5872 be used for mass spectrometry?
Yes, our Protease/Phosphatase Inhibitor Cocktail (100X) #5872 can be used in mass spectrometry (MS) experiments. This cocktail does not contain AEBSF, which can cause peaks to shif...
How should I search for the Pan-Methyl Lysine PTM in LC-MS/MS data when using your PTMScan® Pan-Methyl Lysine Kit #14809?
Our PTMScan Pan-Methyl Lysine Kit #14809 will enrich for all three forms of methylated lysine: mono, di and tri-methyl lysine. To reduce the complexity of the search space, it is n...