Bio-rad Trans-Blot® Plus Cell User Manual

Trans-Blot®PlusElectrophoreticTransfer CellInstruction ManualCatalog Number170-3990For Technical Service Call Your Local Bio-Rad Office or in the U.S.

2.4 Assembling the Gel Sandwich and Cassette AssemblyEach gel sandwich will contain the gel and membrane sandwiched betweentwo pieces of blot absorbe

Note: Extra care is required when handling large gels, first align one side of the gelwith the side of the filter paper and slowly lower the rest of t

Note: The color-coded cables on the lid MUST attach to the electrode cardsof the same color. Reversing the orientation of the cables will cause irrev

Section 3Transfer Conditions3.1 General Guidelines and Running ConditionsThe electric field strength (V/cm) is the driving force in electrophoretic t

Table 3.1 SDS-PAGE GelsThese conditions were determined empirically using 12.5% Tris-HCl Criterion gelsand prestained SDS-PAGE molecular weight stand

Table 3.2 Native GelsThese conditions were determined empirically using 12.5% Tris-HCl Criteriongels and native horse myoglobin samples. See Section

Table 3.3 Isoelectric Focusing, Native, Acid Urea Gels, BasicProteinsThese conditions were determined empirically using 12.5% Tris-HCl Criteriongels

Table 3.4 DNA and RNAThese conditions were determined empirically using 5% uniform TBE Criteriongels and the low range Fluorescein labeled DNA stand

2. Current limits The PowerPac 200 Power Supply is capable of a 200 Volt output. Unless acurrent limit is set, uncontrolled conductivity changes may r

10. These variables will change total resistance and current readings:• Alterations to the buffer make-up, (e.g., addition of SDS or changes in ion co

Table of ContentsPageSection 1 General Information ...11.1 Introduction...

Towbin Buffer with 10 % Methanol25 mM Tris, 192 mM glycine, 10% v/v methanol, pH 8.3a) Using 10X Tris /glycine buffer (catalog #161-0734 for 1L bottle

CAPS BufferCAPS-based transfer buffers (10 mM CAPS, 10% methanol, pH 11) may bepreferable for transfers of high molecular weight proteins (e.g. >50

Dunn carbonate bufferIn some cases, this buffer may produce higher efficiency transfers and improvethe ability of antibodies to recognize and bind to

TAE (Tris-Acetate EDTA)40 mM Tris-Acetate 1 mM EDTAa) Using 50X TAE buffer (catalog #161-0743 for 1L bottles or catalog # 161-0773 for 5L cube) 20 ml

4. Vary buffer type and pH.a. Reduce the buffer strength. Dilution of transfer buffer results in lower current at any given voltage. This will allow t

4.2 Optimizing DNA and RNA TransferAltering the gel percentage can solve problems with elution of nucleic acids. Itmay be somewhat more difficult to

PVDF MembraneBio-Rad offers PVDF (Polyvinylidene difluoride) membranes that are ideal forimmunoassays of blotted proteins (Immun-Blot PVDF) or amino-t

Section 6Troubleshooting GuidePoor transfer of proteins1. Transfer apparatus is assembled incorrectly and the proteins are moving in thewrong directio

Swirls or missing bands; diffuse transfers1. Poor contact between the membrane and the gel. Air bubbles or excess bufferremain between the blot and ge

• Use Tween-20 detergent in the wash and antibody incubation steps.Reduce or eliminate the more stringent washing conditions.4. SDS in the transfer bu

Section 1General Information1.1 IntroductionThe Trans-Blot Plus cell is an electrophoretic transfer cell designed for use withlarge format gels, such

Section 7MaintenanceCleaning• After transfer, remove at least half the buffer remaining in the tank beforeattempting to lift or move the tank from the

Section 8Product InformationCatalogNumber Product description170-3990 Trans-Blot Plus Cell with Plate Electrodes and Super CoolingCoil, includes 3 ge

Section 9References1. Burnette, W. N., Anal. Biochem., 112, 195 (1981)2. Erickson, P. G., Minier, L. N. and Lasher, P. S., J. Immun. Meth., 51, 241 (1

Section 10WarrantyThe Trans-Blot Plus cell electrophoretic transfer cell is warranted for one (1)year against defects in materials and workmanship. If

Life ScienceGroupWeb sitewww.bio-rad.com Bio-Rad Laboratories Main OfficeAlso in: AustraliaPh. 02 9914 2800, Fx. 02 9914 2889 AustriaBelgium Ph. 09

21.2 SpecificationsTrans-Blot Plus cell tankOverall dimensions 39.4 cm x 17.27 cm x 30 cmMaterial AcrylicBuffer requirement 12 litersBuffer capacity

1.3 Safety Power to the Trans-Blot Plus cell is supplied by an external DC voltage powersupply. This power supply must be ground isolated in such a w

Trans-Blot Plus Cell Assembly of PartsFig. 1. Gel Holder Cassettes (3)Cathode Plate (Black)Super Cooling CoilLidDrain PortHandlesAnode Plate (Red)Buff

Section 2Set up and Basic operation2.1 ComponentsBuffer Tank and LidThe buffer tank and lid combine to fully enclose the inner chamber during electro

2.2 Additional ComponentsMagnetic stir plateA magnetic stir plate with a surface area that is sufficient to accommodate theTrans-Blot Plus cell is re

Fig. 2. 4. If necessary, adjust the position of the cathode leads (black) on the lid so thatthey correspond to the position of the cathode electrode