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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 readout of cytokine treatment is measured in a standardized bioassay. The ED50 (expressed in ng/ml), which is the concentration of cytokine or growth factor required to induce half the maximal biological response, is determined in the assay. The following formula can be used to convert the activity as an ED50 in ng/ml to specific activity in units/mg:
Specific activity (units/mg) = 10E6 / ED50 (ng/ml).
For example, the ED50 for our Human Leukemia Inhibitory Factor (hLIF) #8911 is approximately 0.85ng/ml (based on the median ED50 range from the webpage/datasheet) and therefore the specific activity would be 1.17 X 10^6 Units/mg:
10ug = 11,700 Units
50ug = 58,500 Units

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With adherent cell lines, one first needs to detach the cells from the dish. We recommend using trypsin to detach the cells. Alternatively, you can use accutase, but the resulting cell pellet doesn't pack as well during centrifugation, so you tend to lose cells during the wash steps. Suspension cells are much easier. Just count the cells using a hemacytometer or auto cell counter and then collect the number of cells you need. We don't observe a significant difference in assay performance with adherent versus suspension cells.

Cellular Pathways to Metabolism

Cellular homeostasis is regulated by several key metabolic pathways that maintain energetic status and provide the building blocks for proper function.

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Regrettably, we do not test our Streptavidin-HRP #3999 for use in ELISA . Therefore, we cannot provide any firm guarantees or a recommended dilution. However, it should work for ELISA with optimization. We suggest starting with a 1:2000 dilution (0.1 µg/mL).

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The Human ACE2 (18-652) Recombinant Protein (mFc-Tag) #83986 is not expressed with viral vectors. The gene for the Fc fusion is encoded on a plasmid and expressed by transient expression. There are some viral sequences such as promoters and polyadenylation sequences present in the expression construct. It is extremely unlikely to be contaminated with viral vector sequences such as in retro or lentiviral systems.

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.

T Cell Receptor Signaling

Expert-reviewed interactive pathway providing a current overview of T Cell Receptor (TCR) Signaling.

Mechanisms of Cell Death: Necrosis & Necroptosis

Activated by pro-inflammatory signaling, ischemic injury or viral infection, necroptosis is a programmed regulated form of necrosis requiring RIP3 and MLKL

Cell Process: Seven Assays to Detect Apoptosis

Apoptosis is a highly regulated form of programmed cell death that occurs in multicellular organisms throughout life and in response to cellular stress.

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The average diameter of each Concanavalin A bead particle is ~1.0µm while the size range of a cell is 0.1-100 µm (the most commonly used cell lines like HeLa, HepG2 and HEK293 all have a cell size > 10µm). Therefore, in most cases, a cell is coated by multiple beads. This has also been confirmed by inspection under a microscope.

Restoring a Living Shoreline: CST Volunteers with Salem Sound Coastwatch

As part of our commitment to sustainability, CST employees recently participated in a volunteer day with local nonprofit, Salem Sound Coastwatch (SSCW).

Need Better ChIP Results? Use Enzymatic Digestion

Learn how enzymatic digestion can help you get better ChIP results.

Resources

Both SimpleChIP® ChIP-seq Multiplex Oligos for Illumina® (Single Index Primers) #29580 and SimpleChIP® ChIP-seq Multiplex Oligos for Illumina® (Dual Index Primers) #47538 are compatible with the Illumina NGS system. The difference is that the single index primer can only label up to 12 different samples while dual index primers can label up to 96 different samples. Therefore, if you hope to pool together more than 12 samples as one sequencing sample submission, you should purchase our dual index primers.

Writing a Grant Part 4: Additional Details

A research timeline can demonstrate that you've considered project feasibility and future directions show that your plans span beyond the current proposal.

Regulation of Microtubule Dynamics

Expert-reviewed interactive pathway providing a current overview of Regulation of Microtubule Dynamics.

Translational Control

Expert-reviewed interactive pathway providing a current overview of Translational Control.

Regulation of Actin Dynamics

Expert-reviewed interactive pathway providing a current overview of Regulation of Actin Dynamics.

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The antigen used to produce our Vav1 (D45G3) Rabbit mAb #4657 is 100% conserved in Vav1 isoforms 1, 2 and 3. Therefore, we would expect this antibody to recognize these three isoforms.

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实验室期望-CST的官方博客，我们将讨论您在工作时的期望，我们将分享提示，技巧和信息，以确保您在工作时的时间达到这些期望。

Successful Immunofluorescence: Antibody Dilution & Incubation Conditions

Changing the primary antibody dilution and incubation conditions affects fluorescence intensity and signal-to-noise ratio in IF experiments.

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Boiling lysates for the detection of mGluR5 should be avoided as it can cause loss of signal. The mGluR5 protein is unstable at higher temperatures and boiling of the lysate will cause it to degrade to the point where it can no longer be detected by our mGluR5 (D6E7B) Rabbit mAb #55920.

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Our CUT&RUN Assay Kit #86652 is compatible with downstream qPCR analysis. The protocol with the kit provides instructions and reagents for generating an input DNA sample that can be used to quantify your CUT&RUN DNA by qPCR. For qPCR analysis of CUT&RUN DNA, we recommend designing primers that generate amplicons of 60 to 80 bp in length.

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No, we do not observe any sequence bias in our CUT&RUN assays. Free MNase may have some sequence bias in the genome, however, in CUT&RUN, the MNase is fused with pAG and recruited by the target-specific antibody. The digestion to chromatin is directed to where the antibody locates in the genome. Therefore, the antibody specificity and not the sequence bias of MNase has more of an effect on the result.

The Importance of Research Antibody Specificity and Reproducibility During the SARS-CoV-2 Crisis

While healthcare providers and researchers race treat COVID-19, the quality of the tests themselves threatens to undermine their progress.

Immunoprecipitation Experimental Design Tips

Immunoprecipitation (IP) is a technique used to enrich a specific protein from a heterogeneous cell or tissue extract using a target-specific antibody. Co-immunoprecipitation (co-IP) is the pull-down of intact protein complexes.

When to use Immunohistochemistry (IHC)

Understand when to use immunohistochemistry (IHC) over any other technique including Immunocytochemistry, Western Blot and ELISA

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.

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Glut family proteins, and possibly other membrane-bound targets, can form aggregates when heated and reduced, which can inhibit their ability to migrate in SDS-PAGE. This may result in the observation of larger than expected band sizes, multiple bands, or no signal at all.

Immunohistochemical Analysis

Immunohistochemical Analysis. Learn the best ways to interpret IHC staining and understand experimental results accurately. Click here.