Protein Preparation and AGE Detection by Slot Blot
SpeedBlot (His) is equally suitable for Western Blot assays and Dot or Slot Blot
Bacterial cells underwent lysis, and denatured DNA was fixed on a nylon membrane (GE Healthcare Life Science, Sao Paulo, Brazil) using the checkerboard slot blot
device (Minislot 30, Immunetics, Cambridge, USA).
Previous methods of quantifying hBDs in body fluids involved acid extraction followed by slot blot
assays (14,15), semiquantitative Western analysis (16-18), or RIA (19).
Total hsp70 isoforms in oyster gills were detected with a monoclonal antibody raised against human hsp70 (Affinity Bioreagent, 3A3) and total hsp70 quantified using the slot blot
assay (Cruz-Rodriguez & Chu 2002, Lewis et al.
analysis.--Prepared samples (20 [micro]g total protein) were blotted onto PVDF membranes (Micron Separations, Westboro, MA, USA) using a Rio-Dot SF apparatus (BioRad).
(13) Not to be constrained by geometry, the material can also be applied as a line, and the process is then referred to as a slot blot
. If the probe is immobilized and the amplified target is labeled, the assay is then referred to as a reverse dot blot or a "blot dot." (14) These methods are well suited to high-throughput needs, as many samples can be spotted and probed in a single reaction in the dot-blot scenario, and a sample can be reacted with many probes in one reaction in a blot-dot paradigm.
Several technologies are used in this field: Southern blot hybridization, dot blot and slot blot
hybridization, Northern blot hybridization, PCR, RT-PCR, among other types of amplification reactions.
Number of Tests Cost/Test Product Method per Kit ($) Digene In situ 20 15 Enzo In situ 40 10 ONCOR Southern blot 60 7 Virapap Slot blot
50 7 ViraType In situ 20 15 learn.
This study reports the development of an adaptation for the RPA which allows accurate quantitative analysis of large numbers of mRNA samples by replacing PAGE with direct application of the digested sample onto the hybridization membrane by way of a slot blot
. To test this protocol, three experiments of three runs each were performed using dilution series of 6 to 200 pg of interleukin-2 sense RNA transcript.