Slot Blot Apparatus
- S And S Mini Fold Slot Blot Apparatus
- Minifold I Slot-Blot Replacement Plate, 48 Wells 1/pk
- Blotting Apparatus: MiniBlotter, MiniSlot, Western Blot Box ...
A dot blot (or slot blot) is a technique in molecular biology used to detect proteins. It represents a simplification of the western blot method, with the exception that the proteins to be detected are not first separated by electrophoresis. Instead, the sample is applied directly on a membrane in a single spot, and the blotting procedure is performed.
Each dot or slot blot would contain known amounts of target protein or cell lysate. Once dry, dot blots and slot blots are subjected to the same immunodetection steps used for Western blotting, i.e. Blocking, antibody incubation, and target detection with substrate. Typical dot blot membrane. Darker dots indicate more protein. A dot blot (or slot blot) is a technique in molecular biology used to detect proteins. It represents a simplification of the western blot method, with the exception that the proteins to be detected are not first separated by electrophoresis. The 96-well Bio-Dot ® and 48-well Bio-Dot SF (slot format) microfiltration units provide easy, reproducible methods for binding proteins or nucleic acids in solution onto membranes. Many experimental protocols can be accommodated by using interchangeable templates. Bio-Rad dot blot apparatus Dot Blot Apparatus, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 255 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more. Description Four different sample number, size and style of Hybridisation Manifold are offered; two types of Dot Blotter and two types of Slot Blotter. Typical applications include clone screening with DNA/RNA probes in Southern/Northern blots and immunological screening with antibodies in Western blots.
The technique offers significant savings in time, as chromatography or gel electrophoresis, and the complex blotting procedures for the gel are not required. However, it offers no information on the size of the target protein.[1]
Uses[edit]
Performing a dot blot is similar in idea to performing a western blot, with the advantage of faster speed and lower cost.
Dot blots are also performed to screen the binding capabilities of an antibody.[2]
Methods[edit]
A general dot blot protocol involves spotting 1–2 microliters of a samples onto a nitrocellulose or PVDF membrane and letting it air dry. Samples can be in the form of tissue culture supernatants, blood serum, cell extracts, or other preparations.[3]
The membrane is incubated in blocking buffer to prevent non-specific binding. It is then incubated with a primary antibody followed by a detection antibody or a primary antibody conjugated to a detection molecule (commonly HRP or alkaline phosphatase). After antibody binding, the membrane is incubated with a chemiluminescent substrate and imaged.
Apparatus[edit]
Dot blot is conventionally performed on a piece of nitrocellulose membrane or PVDF membrane. After the protein samples are spotted onto the membrane, the membrane is placed in a plastic container and sequentially incubated in blocking buffer, antibody solutions, or rinsing buffer on shaker. Finally, for chemiluminescence imaging, the piece of membrane need to be wrapped in a transparent plastic film filled with enzyme substrate.
Vacuum-assisted dot blot apparatus has been used to facilitate the rinsing and incubating process by using vacuum to extract the solution from underneath the membrane, which is assembled in between several layers of plates to ensure good seal between sample wells, hold waste solution, and deliver suction force. For chemiluminescence signal detection, apparatus need to be disassembled and the membrane need to be taken out and wrapped in a transparent plastic film.
See also[edit]
References[edit]
S And S Mini Fold Slot Blot Apparatus
- ^'Test Blots, Slot Blots & Dot Blots - Immunodetection Bio-Rad'. Bio-Rad.
- ^Rupprecht, Kevin R.; Nair, Rad K.; Harwick, Larissa C.; Grote, Jonathan; Beligere, Gangamani S.; Rege, Sushil D.; Chen, Yon-Yih; Lin, Zhen; Fishpaugh, Jeffrey R. (15 December 2010). 'Development of a dot-blot assay for screening monoclonal antibodies to low-molecular-mass drugs'. Analytical Biochemistry: 160–164. doi:10.1016/j.ab.2010.08.003. PMID20696124.
- ^'Dot Blot Protocol'(PDF).
Minifold I Slot-Blot Replacement Plate, 48 Wells 1/pk
