Technical Support Articles
Results (713)
How many LC-MS/MS injections are obtained from a PTMScan® enrichment?
Each PTMScan® enrichment contains sufficient peptides for three to four LC-MS/MS runs. Therefore, one enrichment can produce enough peptides for technical replicates and enough rem...
Why does MAFA (D2Z6N) Rabbit mAb #79737 recognize a band at approximately 48 kDa when the predicted molecular weight is approximately 37 kDa?
MAFA is a phosphoprotein. The multiple phosphorylation sites on this protein cause a mobility shift, resulting in the protein migrating slower by gel electrophoresis. This has been...
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. ...
Why am I seeing dark background all over my western blot when using your Phospho-p53 (Ser15) Antibody #9284?
It is critical that our Phospho-p53 (Ser15) Antibody #9284 be diluted into 5% w/v non-fat dry milk, 1X TBS, 0.1% Tween® 20 and incubated at 4°C with gentle shaking, over...
What DNA library prep kit do you recommend using for the CUT&Tag assay?
We recommend using our CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415 to make...
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 ...
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...
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...
Can the dot blot protocol be modified if I don't have a dot blotting apparatus for use with your 5-Methylcytosine (5-mC) (D3S2Z) Rabbit mAb #28692?
If a dot blotting apparatus is not available for use, we recommend denaturing your DNA in a lower volume/higher concentration (see modified steps below):
Section B. Dot Blot Mod...
What secondary antibodies do you recommend for fluorescent western blotting?
We currently have four different secondary antibodies approved for use in fluorescent western blotting, depending on species and wavelength of detection:
- Anti-mouse IgG (H+...
Can you help me calculate the bioactivity of my cytokine?
Specific activity (expressed in units/mg) is a function of cytokine or growth factor purity and appropriate protein folding. To determine the specific activity, typically the reado...
Which beta-actin antibody do you suggest for paraffin-embedded immunohistochemistry (IHC-P) with mouse samples?
We have several antibodies for β-actin that are validated for IHC-P in mouse samples. These are β-Actin (13E5) Rabbit mAb #4970 and β-Actin (8H10D10) Mouse mAb #3700. We would reco...
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...
What are the high molecular weight bands detected by the β1-Adrenergic Receptor Antibody #12271?
The antigen for the β1-Adrenergic Receptor Antibody #12271 surrounds Gly406 of the human β1-Adrenergic Receptor protein and is contained entirely in the cytoplasmic domain. #12271 ...
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 formulat...
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...
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 #68...
Will your GFP (D5.1) Rabbit mAb #2956 detect EGFP, eBFP2, and/or mCerulean3?
Our GFP (D5.1) Rabbit mAb #2956 is expected to detect EGFP, eBFP2, and mCerulean3.
Does Cell Lysis Buffer (10X) #9803 work with the Bradford assay?
Cell Lysis Buffer (10X) #9803 will interfere with the Bradford protein assay. This is because it contains detergent. To make the assay somewhat compatible, dilute your protein quan...
What procedure do you recommend for antigen retrieval with EDTA?
We recommend using a microwave to bring slides to a boil in 1 mM EDTA, pH 8.0 followed by 15 minutes at sub-boiling temperature. Typically the power level must be adjusted during t...
Are the Protein A Magnetic Beads #73778 and the Protein G Magnetic Beads #70024 hydrophilic or hydrophobic?
The base particle of the Protein A Magnetic Beads #73778 and the Protein G Magnetic Beads #70024 is somewhat hydrophilic. It is difficult to predict the hydrophilicity or hydrophob...
Can I use PFA as a fixative rather than the fixative provided with the Senescence β-Galactosidase Staining Kit #9860?
We have not tested samples that have been fixed with paraformaldehyde so we cannot guarantee the kit will work using this fixative. For reference, our Fixative Solution #11674 whic...
Can I freeze the PTMScan® HS Magnetic Immunoaffinity Beads?
The PTMScan HS Magnetic Immunoaffinity Beads should not be frozen. We recommend storing this product at 4C. However, we have performed functional testing by IAP and subsequent mass...
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 ...
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.
What if my yield of CUT&Tag DNA is too low to see on an Agilent Bioanalyzer or TapeStation system?
Due to the expected and typically low yield of CUT&Tag DNA, we recommend using all 30 µL of the CUT&Tag DNA sample (from Our Tau (D1M9X) XP® Rabbit mAb #46687 is predicted to recognize 2N4R (tau40), 2N3R (tau39), 1N4R (tau46), 1N3R (tau37), 0N4R (tau24), and 0N3R (tau23), as all six isoforms include ... Yes. Cell fixation is recommended when the cells are very fragile (easily lysed during experiments), when the cellular signaling pathway is sensitive to the Concanavalin A beads... We prepare Jurkat -/+ Anti-CD3 antibody (10 µg/mL for 2 min) cell extracts as follows:
Does your Tau (D1M9X) XP® Rabbit mAb #46687 recognize all six Tau isoforms?
Is it possible to perform CUT&Tag on fixed frozen cells?
Can you share your protocol for preparing Jurkat -/+ Anti-CD3 antibody (10 µg/mL for 2 min) cell extracts?
Which Cadherins does the Pan-Cadherin Antibody #4068 detect?
The Pan-Cadherin Antibody #4068 can detect R-, N-, E-, and VE-Cadherin. It does not detect P- or K-Cadherin. Reactivity to other Cadherins has not been tested.