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Technical Support Articles

Results (709)

Can I use your Phosphate Buffered Saline with Tween® 20 (PBST-20X) #9809 to make my primary, blocking, and secondary buffers for western blotting?

Our in-house testing indicates that using Phosphate Buffered Saline with Tween® 20 (PBST-20X) #9809 as the base buffer for washes, blocking, and antibody incubation can interfere w...

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 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 ...

Can I use DNA affinity columns to purify CUT&RUN DNA for transcription factors?

Yes, we recommend using our 

The DNA yield for my CUT&RUN experiment seems a bit low compared to what I was getting in my ChIP experiments. How can I improve the yield?

A ChIP experiment uses 4M cells per reaction while a CUT&RUN assay only uses 5K to 100K starting cells, so it is expected to see a lower DNA yield from a CUT&RUN assay than...

Is the Phosphate Buffered Saline (PBS-1X) pH7.2 (Sterile) #9872 RNase free?

Our Phosphate Buffered Saline (PBS-1X) pH7.2 (Sterile) #9872 (both L and S sizes) is tested for sterility and endotoxin levels. All new lots are subjected to a 2 week sterility tes...

Will particulates in the protein substrate tubes of the GTPase kits negatively affect the results of the assay?

It is not uncommon to see particulates in the protein substrate tubes provided with the GTPase kits. After extensive testing, we have found that these are not a cause for concern. ...

Are the Protein G Agarose Beads #37478 compatible with goat antibodies?

While we have not tested our Protein G Agarose Beads #37478 with goat antibodies in-house, goat IgG has a strong binding affinity to protein G. Therefore, our Protein G Agarose Bea...

Are your siRNAs pools or single duplexes?

All of our siRNAs are single duplexes. We discontinued pools a number of years ago to avoid off-target effects.

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 

Why are there particulates in my sample of Animal-Free Blocking Solution (5X) #15019?

Particulates may occasionally be seen in our Animal-Free Blocking Solution (5X) #15019 due to material becoming caught in the threads of the cap/bottle and drying out over a short ...

Why does the material in my new vial of Echinomycin #51434 look different?

Echinomycin #51434 is lyophilized into the vial, and any difference in the appearance of the lyophilized echinomycin is due to a dry coating of the compound in the vial. When you a...

Are the -20C components within the CUT&RUN Assay Kit #86652 stable to use if they have been stored at 4C for a period of time?

Upon arrival of the kit to your facility, the components should be broken down and stored at their recommended temperatures. We have conducted stability tests for a few components ...

To store plates long-term when using the Senescence β-Galactosidase Staining Kit #9860, do I need to wash off the β-Galactosidase staining solution prior to overlaying with 70% glycerol?

In section C, part 8 of the Senescence β-Galactosidase Staining Kit #9860 protocol, you may overlay the plates with 70% glycerol directly after removing the staining solution. It i...

Do you recommend performing size selection of the DNA during library preparation for CUT&RUN or ChIP-seq?

We do not recommend performing size selection of the DNA during library preparation for CUT&RUN or ChIP-seq. Based on our experience and feedback from customers, size selection...

Are your Anti-rabbit IgG, HRP-linked Antibody #7074 and Anti-mouse IgG, HRP-linked Antibody #7076 polyclonal or monoclonal?

Our Anti-rabbit IgG, HRP-linked Antibody #7074 and Anti-mouse IgG, HRP-linked Antibody #7076 are both polyclonal products.

Do you have any tips for using FACS-isolated cells in CUT&RUN experiments?

We have no specific tips for using FACS-isolated cells in CUT&RUN experiments. Like a cultured cell suspension, if your isolated cells are in less than 40ul PBS, you can move d...

How do I identify O-GlcNAc peptides by mass spectrometry after PTMScan®?

The PTMScan O-GlcNAc [GlcNAc-S/T] Motif Kit #95220 employs an antibody that will immunoprecipitate peptides with the native O-GlcNAc PTM on serine (S) or threonine (T) residues. To...

Does your CD44 (156-3C11) Mouse mAb #3570 recognize all variants of human CD44?

The CD44 gene encodes 20 exons. Alternative splicing gives rise to CD44s and multiple CD44 variants (CD44v). CD44s is encoded by constant exons 1-5, 16-18, and 20. The CD44 vari...

Does the performance of your Human ACE2 (18-615) Recombinant Protein (hFc-Tag) #38365 and Human ACE2 (18-652) Recombinant Protein (hFc-Tag) #99339 differ due to the expression system?

At this time, we have no data that suggests the expression platform matters for protein performance. Typically, the difference between expression systems is yield rather than perfo...

What is the percentage of SDS in the ChIP Sonication Nuclear Lysis Buffer #28778?

The ChIP Sonication Nuclear Lysis Buffer #28778 included in the SimpleChIP® Sonication Cell and Nuclear Lysis Buffers #81804 contains 0.1% SDS.

Why are there background peptides in my PTMScan® experiment?

The background or number of non-PTM peptides in each enrichment is primarily due to non-specific interactions with the beads the antibody is conjugated to. Therefore, this backgrou...

What is the composition of your Lysis/Binding/Wash Buffer #11524?

The composition of our Lysis/Binding/Wash Buffer #11524 is as follows: 25 mM Tris-HCl, pH 7.2, 150 mM NaCl, 5 mM MgCl2, 1% NP-40 and 5% glycerol.

Is prewashing the Protein A Magnetic Beads #73778 and Protein G Magnetic Beads #70024 necessary for my IP experiment?

We recommend washing the Protein A and Protein G magnetic beads prior to use to help clear them of any potential increase in background due to the buffer in which they are provided...

Do you have a peptide blocking protocol for IHC?

For IHC, we recommend combining the peptide and the primary antibody at a 10:1 ratio (µg : µg) in the antibody diluent. Incubate the peptide/antibody mixture for a minimum of 30 mi...

Do you observe any differences in CUT&RUN assay performance among different cell lines?

We have performed CUT&RUN on dozens of cultured adherent and suspension cell lines, both human and mouse. So far we have not noticed any significant difference in performance b...

Why don't the phosphorylation sites given in the names of your phospho-EGFR antibodies match the same residue numbers in the UniProt sequence (P00533)?

When EGFR was first reported, the phosphorylation sites and corresponding amino acid numbering did not account for a cleaved signal peptide measuring 24 amino acids in length. Cons...

How are your antibody concentrations determined?

We determine the concentration of our antibodies by measuring the absorbance at 280nm (OD280).

Why am I unable to see phalloidin staining in my methanol-fixed and permeabilized samples?

Phalloidin is only able to bind the native quaternary structure of F-actin. In order to retain the quaternary protein structure, formaldehyde fixation should be utilized. Methanol ...

How do I validate the PTM sites identified in PTMScan® experiments?

The validation of PTM sites may be accomplished in several ways. Often, the site of interest may be seen in the context of more than one peptide due to alternative forms caused by ...

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