How To Decide ELISA Dilution Ratio?

Dilution ratio describes a simple dilution – a unit volume of solute (or sample) is combined with a desired unit volume of solvent (or diluent), to reach a desired total volume (Vsolute + Vsolvent = Total Vsolution)

Thus, a dilution ratio of 1:4 describes 1 part solute + 4 parts solvent = 5 parts total. The sum of both solute plus solvent equals total, final volume.

What Is DAPI?

DAPI binds to and ‘stains’ double-stranded DNA, preferably binding to A-T-rich regions in DNA. DAPI stain is excited by ultraviolet (UV) light, with its largest excitation wavelength at ~360nm, and it produces a vibrant blue color with its largest emission wavelength at ~460nm when bound to DNA. Due to its fluorescent properties and rich blue color, it is readily used for visualization in fluorescence microscopy and other assays

Learn About Recombinant Protein Production

Proteins are the workhorse molecules that drive virtually every biological system. With the increasing recognition of the role of proteins in various research and manufacturing activities, simply isolating them from their natural host cells cannot meet the escalating demand of the market. Chemical synthesis is also not a viable option for this endeavor due to the size and complexity of proteins. Instead, the advances made possible by recombinant DNA technology have profound implications in increasing the supply of proteins needed by the rising demand.

What is CCK-8?

Cell Counting Kit-8 (CCK-8) is a readily available, easy-to-use cell viability and cytotoxicity assay. CCK-8 takes advantage of WST-8 [2- (2- methoxy-4-nitrophenyl)-3- (4-nitrophenyl)-5- (2, 4-disulfophenyl)-2H- tetrazolium, monosodium salt] to directly measure cell viability. Specifically, WST-8 is a water-soluble salt that can be reduced by dehydrogenases in viable cells to a water-soluble formazan dye. The biochemical reaction causes a change in color that can be easily quantified by measuring absorbance at 450nm. This simple and convenient colorimetric assay has been widely used to determine cell viability and cytotoxicity of various agents in cell culture.

Nitrocellulose Membrane For Western Blot

Nitrocellulose membranes are one of the top matrices used in protein blotting. They have high protein-binding affinity, compatibility with a variety of detection methods, and the ability to immobilize proteins, glycoproteins, or nucleic acids. Examples of compatible detection methods include chemiluminescence, chromogenic, and fluorescence. It is proven to produce excellent signal-to-noise results when used for amino acid analysis and western, northern, and Southern blotting.

Predicting Western Blot Band Sizes

First described in 1979, the technique of western blotting has since become one of the most commonly used analytical methods in life science research. Just last week, we received a few questions from confused researchers about weird band sizes in their western blot results:

Why does my result show a different band size than the predicted size?”

“What is the expected band size? Why is it different than the observed band size?”

Which ELISA Is For You?

Are you familiar with the multiple methods you could use to perform an ELISA? Among the standard assay formats illustrated below, where differences in both capture and detection are in concern, it is important to differentiate between the particular strategies that exist specifically for the detection step. However an antigen is captured to the plate (by direct adsorption to the surface or through a pre-coated "capture" antibody, as in a sandwich ELISA), it is the detection step (as either direct or indirect detection) that largely determines the sensitivity of an ELISA.

Guidelines For Preparing ELISA Standards

ELISA (enzyme-linked immunosorbent assay) is a plate-based assay used to detect the concentration of a specific protein in a liquid sample. A purified protein should be used to prepare the standard curve. Otherwise, use a recombinant protein which can be semi-purified in the lab and measure the concentration with HPLC. Some companies may provide purified antigens. Or, save time by purchasing Boster Bio’s PicoKine ELISA kits, which include the standard and other necessary ELISA reagents for your convenience.

5 Pitfalls To Avoid For ELISA

ELISA (enzyme-linked immunosorbent assay) is a convenient and simple method to quantitatively or qualitatively detect peptides, proteins, antibodies, and hormones in samples, rendering it as one of the most widely used immunoassays. Despite the many advantages of conducting ELISA, there are some mistakes that could turn your ELISA experiment sour. Help prevent this situation from happening by avoiding 5 common pitfalls when performing an ELISA:

How To Use Checkerboard Titration To Optimize Your ELISA Immunoassays

In order to get the best results from your ELISA assay, the dilution factors of the sample and the detection antibodies must be optimized. If your sample or antibodies are too concentrated, you risk saturating the assay. If they are not concentrated enough, your signal will be weak and difficult to detect. For strong, quantifiable signal, use a checkerboard titration to test for the optimal concentration of sample and detection antibodies.

How To Perfect Your ELISA Standards

At Boster, one common question we get from researchers is, “How do I prepare the ELISA standard?” We’re glad you asked because proper construction of the standard curve is the very first step for every ELISA experiment. The standard curve can help confirm that the quality of the kit and the operation procedures are acceptable for further steps.

3 Main Types Of IHC/ICC Fixatives

Sample fixation is a required and crucial step for every successful IHC/ICC experiment. Appropriate fixation of samples provides the following benefits during the tissue preparation process.

Choosing which fixing solution to use depends on your sample type and antigen.

6 IHC Controls You Should Know

Immunohistochemistry (IHC) is a popular protein detection method that utilizes antibody-antigen interactions to visualize the distribution and localization of specific cellular components within cells and in their proper tissue context.

In the lab, researchers invest time and effort to optimize the sample preparation and sample staining processes of IHC. When successful, the results produce a strong and specific signal.

5 Tips To Reduce Autofluorescence

The common problem for all fluorescence-based cell detection methods is the cells’ natural fluorescence. Cellular autofluorescence is due to the presence of various biological structures, such as collagen, elastin, NADPH, flavins, mitochondria, and lysosomes, which usually absorb in UV to blue range (355-488 nm) and emit in the blue to green range (350-550 nm). Therefore, autofluorescence interferes with analysis by reducing signal sensitivity and resolution of fluorochromes that operate in that range – FITC, GFP, and Pacific Blue to name a few.