Abnova社動画ページのご紹介

※ 本製品は研究用です。研究用以外には使用できません。

Abnova社が提供する、各種アプリケーションの使用例動画をご紹介します。
Abnova社ホームページでは、下記以外にも多数の動画をご用意しています。詳細はこちらをご覧下さい。

Western Blotting is an analytical technique used to detect specific proteins in a given sample. It uses gel electrophoresis to separate native or denatured proteins by the length of the polypeptide. The proteins are then transferred to a membrane where they are detected using antibodies specific to the target protein.

Dot blot is a technique can be used as a semiqualitative method for rapid screening of a large number of samples. It is for detecting, analyzing, and identifying proteins, similar to the western blot technique but differing in that protein samples are not separated electrophoretically but are spotted through circular templates directly onto the membrane.

Real-time PCR, also called quantitative real time PCR (Q-PCR/qPCR), is used to amplify and simultaneously quantify a targeted DNA molecule. It enables both detection and quantification (as absolute number of copies or relative amount when normalized to DNA input or additional normalizing genes) of one or more specific sequences in a DNA sample.

Reverse transcription PCR includes two steps. The first step is reverse transcription, in which RNA is reverse transcribed to its DNA complement (complementary DNA, or cDNA) using reverse transcriptase and primers. The second step is amplification using traditional or real-time PCR.

Polymerase Chain Reaction (PCR) is a technique to amplify few copies of a piece of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence. There are three major steps (Denaturation, Annealing and Extension) in a PCR and repeat for 30 or 40 cycles.

Primer-BLAST was developed to help users make primers that are specific to the input PCR template. It uses Primer3 to design PCR primers and then submits them to BLAST search against user-selected database. The blast results are then automatically analyzed to avoid primer pairs that can cause amplification of targets other than the input template.

Cre-Lox recombination is a special type of site-specific recombination. The Cre protein is a site-specific DNA recombinase. It can catalyze the recombination of DNA between specific sites in a DNA molecule. These sites, known as loxP sequences, contain specific binding sites for Cre that surround a directional core sequence where recombination can occur. It is often used in the generation of knockout and conditional knockout animals.

Miniprep plasmid DNA extraction kit is used when the starting E. coli culture volume is 1~5 ml of LB broth and the expected DNA yield is 20~30 μg. Other extraction and purification plasmid DNA Kits are available from varying manufacturers, named by size of bacterial culture, includes gigaprep, megaprep, and midiprep

The indirect ELISA is used primarily to determine the strength and/or amount of antibody response in a sample. In the assay, the antigen of interested is immobilized by direct adsorption to the assay plate. Detection of the antigen can then be performed by using a matched set of primary antibody and conjugated secondary antibodies.

In competitive ELISA, unlabeled antibody is incubated in the presence of its antigen. Then these bound antibody/antigen complexes are then added to an antigen coated well. After washing, unbound antibodies are removed. The more analytes in the sample, the less antibodies will be able to bind to antigens in the well. The signal is then detected using labeled secondary antibodies and the decrease in signal is compared to a control. The major advantage of a competitive ELISA is the ability to use crude or impure samples and still selectively bind any antigen that may be present.

The Sandwich ELISA measures the amount of analyte between capture antibody and detection antibody. The analyte needs to have two different epitope sites available for antibody binding.

Immunohistochemistry is a method of detecting the presence of specific proteins in cells or tissues. It is widely used in basic research to understand the distribution and localization of biomarkers and differentially expressed proteins in different parts of a biological tissue.

H&E stain is a popular staining method in histology. Its a combination of two dyes: the basic dye (hematoxylin) and the alcohol-based dye (eosin). In an H&E stain you will usually see both eosinophilia and basophilia: the nuclei of cells basophilic (blue), while eosinophilia is typical of cytoplasmic constituents (pink).

Immunofluorescence is a technique to visualize a specific protein or antigen in cells or tissue sections by binding a specific antibody chemically conjugated with a fluorescent dye. We use the indirect immunofluorescence staining to perform cells fixed on slides and examine under a fluorescence microscope.

The AbVideo on CytoQuest™ CR Dos and Don’ts part 2 concentrates on the proper handling of CytoChipNano and Coupler including the insertion and removal of outlet and inlet plugs into and from the Coupler, and the placement of CytoChipNano in the Chip Receptor.

Cells are cultured under précised conditions; A complete protocol on how to thaw, maintain and freeze attached cells is shown in this AbVideo.

This AbVideo introduces the protocol of preparing suspension cell for counting. Hemocytometer or Cell Counting Chamber is frequently used to count cell numbers such as cell density of cultured cells in research labs and blood count and sperm count in clinical labs. For counting of attached cells, cells are trypsinized, mixed with a dye (Trypan Blue), loaded onto a hemocytometer and then counted under a microscope.

A demonstration on the procedure of using MTT assay to assess the viability and the proliferation of regular cells with absorbance detection at 595nm is shown. This colorimetric assay measures the reduction of yellow 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) by mitochondrial succinate dehydrogenase.

Bradford assay is a rapid and accurate method to determine the concentration of protein. The assay is based on the observation that the absorbance maximum for an acidic solution of Coomassie Brilliant Blue G-250 shifts from 465 nm to 595 nm when binding to protein occurs. Both hydrophobic and ionic interactions stabilize the anionic form of the dye, causing a visible color change. Within the linear range of the assay (~5-25 mg/ml), the more protein present, the more Coomassie binds.