Resources results for "LOX"

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Fibrosis

Products and Related Resources for Fibrosis SARS-CoV-2 Research

COVID-19 and Fibrosis: Long-term damage beyond acute respiratory symptoms

It remains difficult for scientists to predict the long-term health effects of COVID-19, particularly considering its diversity of effects during acute infection. The most severe impacts appear to manifest in tissues with high levels of vascularization (e.g., lungs, heart, kidney, liver), and it is evident that the virus elicits a significant inflammatory response in infected tissues

Safeguard the Quality of Your ELISA Data: Ensuring ELISA Lot-to-Lot Reproducibility

Testing ELISA kit lots for percent coefficient of variation (CV), signal/blank ratios, and optical density helps ensure scientific reproducibility.

The CST Product Performance Guarantee

The guarantee behind all of CST's antibody products, ensuring performance, lot-to-lot consistency, and application validation.

Bulk Quantities and Lot Reservations

Tailor orders to have product when you need it. Make single-lot reservations and order bulk quantities of CST® antibodies, buffers, and ELISA kits.

Resources

There may be some lot-to-lot variability in the signal strength of our Vimentin (V9) Mouse mAb #49636 when testing by western blot due to this antibody being sold by weight instead of reactivity.

What is a Recombinant Antibody and Why is it Important?

Recombinant antibodies offer several key advantages: superior lot-to-lot consistency, continuous supply, and amenability to antibody engineering.

CUT&Tag DNA Library Yield: What to Do if Yours Is Too Low to Detect with an Agilent Bioanalyzer or TapeStation System

CUT&Tag delivers quality NGS data even if the signal from the Bioanalyzer or TapeStation System is low and the DNA library yield is low.

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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 performance is often obtained at a lower concentration, maintaining strong target signal while minimizing background. As such, the concentration of our antibodies varies between products and sometimes between lots. We perform extensive lot-to-lot validation to ensure consistent performance and signal intensity.

Immunoprecipitation Troubleshooting Guide

Immunoprecipitation Troubleshooting Guide for easy to solve high and low background issues.

Western Blotting Troubleshooting Guide

Western Blotting troubleshooting guide for easy to solve high and low background issues.

Resources

The concentration of our Digitonin Solution #16359 is proprietary. The concentration will vary between lots, as we internally test each lot to match performance before release. This is due to activity changing between each resuspension of digitonin used to generate a new lot. This is why we match the efficiency rather than concentration between lots. Therefore we recommend titrating by volume to determine optimal effectiveness. If the cells are very sensitive to digitonin, we recommend starting with 2.5ul digitonin solution per ml buffer and 3-5 titration points. Our experience indicates that adding a higher digitonin amount than required does not significantly affect the result. If the cell line is very insensitive to digitonin, we recommend titrating within the range of 25-100ul digitonin solution per ml buffer.

Hallmarks of Validation - Ranged Strategy

A ranged validation strategy includes both endogenous and heterologous models that express high, moderate, and low levels of the target of interest.

Hallmarks of Cancer: Deregulating Cellular Energetics

Cancer cells need a lot of energy to grow fast—to do so, they show abnormal metabolic pathways.

Fluorescent Staining Using Multiple Antibodies: Two Common Techniques

Co-staining with multiple antibodies is a low-content form of multiplex analysis that be achieved using different host species or fluorophore conjugates.

Hypoxia and Its Role in Metabolism

Hypoxia causes cellular stress and alters normal metabolic activity. These pathways allow cells to survive and recover during bouts of low oxygen.

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Some CUT&RUN protocols utilize contaminating E. coli DNA in the pAG-MNase for sample normalization. However, this can become problematic when switching lots of pAG-MNase, because each lot of enzyme will have differing amounts of contaminating DNA. Instead, we recommend using a separate yeast spike-in DNA sample. We provide a yeast spike-in DNA sample with our CUT&RUN Assay Kit #86652 and our CUT&RUN pAG-MNase and Spike-In DNA #40366 for which every lot is tested and optimized for CUT&RUN. This yeast spike-in DNA provides better lot-to-lot consistency for your normalization efforts.

Hallmarks of Antibody Validation: Heterologous Strategy

Validation using recombinant proteins or exogenous expression in a surrogate cell line can be used for targets with low levels of protein expression.

Resources

Unfortunately, due to the nature of lot production, none of the components within the Senescence β-Galactosidase Staining Kit #9860 are available for individual purchase outside of the kit.

Beat the Blues of Writing a Scientific Rebuttal

Publishing your first “first” author manuscript can be a nightmare, but it taught me a lot about maintaining diplomacy and keeping calm while responding to reviewers.

CUT&RUN Resource Center

Learn about CUT&RUN - a low cell number alternative to ChIP-qPCR and ChIP-seq to analyze protein-DNA interaction in chromatin.

Hallmarks of Antibody Validation: Heterologous Strategy

For antigenic targets where expression of the protein is very low or unknown, the use of recombinant proteins or exogenous expression in a surrogate cell line may be necessary for antibody validation.

Five Reasons to Choose a Conjugation Service Over an Antibody Labeling Kit

DIY conjugation kits let researchers label an antibody themselves but lengthy protocols, poor conjugate quality and low yield cause inconsistent results.

Resources

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 Section VI of the CUT&Tag protocol #77552) for library amplification. While CUT&Tag DNA libraries generated for histone modifications typically show some signal on the Agilent Bioanalyzer or TapeStation systems, libraries generated for non-histone proteins such as transcription factors and cofactors often have very weak or even no visible signal on Bioanalyzer or TapeStation systems, but they still generate NGS results with high-mapping rates, high numbers of identified binding peaks, and decent signal-to-noise ratios across the whole genome. For CUT&Tag libraries where the Bioanalyzer or TapeStation system is unable to identify the average size of the library, we suggest using a size o…

Custom ELISA Kits and Matched Antibody Pairs

Choose custom CST® ELISA products to match your project needs. Reserve lots, customize volumes or kit formats, order custom-matched antibody pairs, and more.

FastScan™ ELISA (Enzyme-Linked Immunosorbent Assay) Kits

FastScan ELISA kits detect key signaling proteins using a solution-based assay format and are validated to ensure reproducible results across lots and users.

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For a recently purchased antibody, the concentration will be found either printed directly on the vial label or in the Certificate of Analysis (CoA). CoAs for the current shipping lot of many of our products are available on the right-hand side of the product webpage next to the data images.

For the concentration of a previous antibody lot, please contact CST technical support by submitting a form online or emailing us at [email protected].

Resources

Yes, there are a lot of tips for CUT&RUN DNA library preparation, especially for CUT&RUN assays with an extremely low number of cells. Compared to ChIP-DNA samples, CUT&RUN DNA has two unique features. First, it is smaller in size and, secondly, the yield of DNA is much lower due to fewer starting cells being used in a CUT&RUN assay.

Because CUT&RUN DNA is small, you need to decrease the incubation temperature from 65C to 50C during end prep and A tailing to prevent the generation of ssDNA. Also, because of its smaller size, you may benefit from using 1.1X volumes instead of 0.9X volumes of Ampure beads during library DNA purification. Importantly, the extension time during PCR amplification of the library can be decreased from 75s to 10-15s. This not only saves time during the PCR program, but it also excludes the possibility of large non-specific DNA being amplified.

Since CUT&RUN DNA has a low yield, you will also need to add less adaptor DNA for ligation in or…

The Utility of Epitope Tags in Western Blot and Immunoprecipitation

Watch this video to learn how epitope tags can be leveraged in your experiments when your protein of interest is newly discovered, expressed at low abundance, or poorly immunogenic.

Resources

If you start with 100,000 cells, you typically get 0.6-6ng DNA. However, if you start with 5,000 - 10,000 cells, the DNA yield can be as low as 0.1-0.2 ng or even undetectable by the picogreen assay. In this case, we suggest you directly proceed with library prep using 20 PCR amplification cycles, which will result in a library DNA with a concentration of around 10-30 ng/ul. QC of the CUT&RUN DNA by qPCR can be performed on the library DNA if you hope to save as much CUT&RUN DNA as possible for the library prep.