Molecular weight marker for Western blotting
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SERVA Western blotting
SERVA VisiBlot Standard I is a ready-to-use mixture of recombinant proteins in the molecular weight range from 25 kDa to 150 kDa. For control during electrophoresis runs and for transfer control after blotting, three protein bands are pre-stained. The remaining seven unstained proteins have IgG binding sites. These are detectable by antibody detection on the blot membrane and allow the exact assignment of the molecular weight.
Popular standard markers for SDS PAGE and Western blotting are also the SERVA Triple Color Protein Standard II and the SERVA Triple Color Protein Standard III.
SDS PAGE gels for Western blotting
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SERVAGel™ TG PRiME™ mini gels
High resolution, efficient transfer and a sensitive detection system are the most important requirements for best results in Western blot experiments. With the use of SERVAGel™ TG PRiME™ ready-to-use gels, excellent separation of your samples is achieved. More information about the gels and the matching mini electrophoresis chamber can be found here: BlueVertical™ PRiME™
SERVAGel™ Neutral HSE mini gels
SERVAGel™ Neutral HSE is optimised for short run times. By using a lower acrylamide concentration, the run time is minimised to 20 minutes. No special solutions or equipment are needed. Another plus is the long shelf life of 15 months. More information about the gels and the matching mini electrophoresis chamber can be found here: BlueVertical™ PRiME™
2DHPE™ BlotGels
The non-covalently bound flatbed 2DHPE™ BlotGels are the first large-format, blotable, film-supported gels on the market. Perform your 2D electrophoresis as usual. After cutting a small strip off gel, you can easily peel the gel off from the film and transfer it to a membrane. More information about the gels and the matching flatbed chamber can be found here: HPE™ BlueHorizon™
Transfer buffer
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Xpress Blotting kits
In addition to the Towbin buffer for small proteins and the Semi-Dry Blotting Buffer Kit (discontinuous buffer system), we have developed various Xpress blotting kits for you. Each kit contains ready-to-use buffers and blotting fleece (80mm x 85 mm). On the one hand, the buffer reduces the transfer time to 15 minutes, and on the other hand, the newly developed blotting fleeces diminish interferences during transfer.
Decide for the Xpress Blotting Kit without membranes to use your own membranes or choose a kit with pre-cut membranes, e.g. Xpress NC Blotting Kit or Xpress PVDF Blotting Kit.
Transfer membranes
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Nitrozellulose membranes
Nitrocellulose membranes are suitable for Western, Southern and Northern blots. The reason for this is the high binding capacity and a low background. Above all, the fleece backing nitrocellulose membranes are a good alternative, since cutting is simple, multiple hybridizations and automatic immobilization are possible.
Polyvinylidenfluorid (PVDF) membranes
PVDF membranes are hydrophobic, have excellent mechanical stability, are highly flexible and tear resistant. In addition, they have high binding capacity and low background. They are also compatible with most staining methods. We therefore recommend Fluorobind membranes to detect proteins with immuno or fluorescence applications.
Blocking reagents
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BlueBlock PF
In order to achieve a good signal-to-noise ratio, it is important to block unspecific antibody binding sites on the transfer membrane.
However, in the case of protein-containing blocking solutions, such as skim milk powder or BSA, there is risk that not only unspecific antibody binding sites are blocked but also specific binding sites are masked. In contrast, BlueBlock PF suppresses only unspecific, while the specific binding sites are unaffected. This increases the signal strength.
Stripping buffer
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BlueClear SB
BlueClear SB is a ready-to-use buffer for efficient stripping of high-affinity antibodies from Western blot membranes after chemiluminescence detection. After incubating the membrane in the stripping buffer for 30-60 min at room temperature, difficult to remove antibodies are efficiently stripped by incubation in the heated buffer. After washing in PBST or TBST, the membrane can be blocked again and incubated with antibodies.
Detection systems
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SERVALight CL HRP substrate kits
SERVALight ready-to-use chemiluminescence substrates detect immobilised proteins (Western blot) or nucleic acids (Southern and Northern blot) either directly with horseradish peroxidase (HRP) or indirectly with HRP-labelled antibodies/streptavidin. The SERVALight substrates are quickly prepared. Just mix solution A (luminol/enhancer solution) and solution B (stabilised peroxide solution) in a 1:1 ratio.
The right one for your application: SERVALight Vega and SERVALight Polaris develop normal light intensity with long duration. SERVALight Eos and SERVALight EosUltra develop high light intensity with a very long duration. SERVALight Helios develops only moderate duration but extremely strong light intensity.
SERVALight PreMix substrate kits
SERVALight PreMix Chemiluminesce Horseradish Peroxidase (HRP) substrates are ready-to-use solutions for convenient and rapid detection of proteins in Western blotting. The premixed solutions save time and increase the consistency of your results by avoiding pipetting errors and possible contamination.
The right one for your application: SERVALight PreMix Vega, SERVALight PreMix Eos, SERVALight PreMix Helios
Detection reagents for Western blotting
SERVAColor BCIP/NBT and SERVAColor TMB blot solutions are ready-to-use and non-toxic. They are used as highly sensitive, colorimetric detections of alkaline phosphatase (AP) or horseradish peroxidase (HRP) in membrane assays. They are characterised by rapid precipitation, low background staining, long stability at room temperature and low fading.
Devices for Western blotting
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BlueVertical™ PRiME™
The BlueVertical™ PRiME™ is a dual mini-tank system with the option of running one or two ready-to-use gels. It can hold gels with external dimensions of 10 x 10 x 0.7 cm. The assembly of the gels is done by a unique clamp system and closes tightly. The easy-to-use blot module turns your chamber into a tank blotter. For more information on the BlueVertical™ PRiME™ system, click here: BlueVertical™ PRiME™
BlueBlot semi-dry blotter
The BlueBlot semi-dry blotter forms a homogeneous electrical field that guarantees fast and efficient transfer of proteins from gel to membrane. The anode is made from platinum-covered steel net, the cathode from stainless steel. Not only the stable acrylic housing is easy to clean but also the long-lasting electrodes can be dismounted from the housing with a flick of the wrist and cleaned under water running tap.
Choose from three sizes. The model BlueBlot SD11 can accomodate one, BlueBlot SD17 four and BlueBlot SD26 six minigels.
BluePower™ power supply for blot applications
The BluePower™ 300 BLOT power supply (300 V, 2 A, 300 W) is suitable for applications requiring high current, such as tank or semi-dry blotting of larger protein gels. Four devices can be connected in parallel and it is freely programmable (9 programmes with 9 steps).
HPE™ BlueHorizon™
The HPE™ BlueHorizon™ is a flatbed system for horizontal electrophoresis with pre-cast gels, self-cast gels and IPG strips. The main applications are isoelectric focusing (IEF), 2D and SDS-PAGE. Two platinum electrodes positionable at three fixed electrode positions allow the use of a wide range of different gel sizes. Blotable, film-supported gels are available. For more information on the HPE™ BlueHorizon™ system, click here: HPE™ BlueHorizon™
Gravity Blotter
The SERVA Gravity Blotter was developed by SERVA for high efficiency blotting of film-supported IEF and SDS PAGE gels. Using press blotting, it is no longer necessary to separate the gel from the support film. The unit consists of a base plate with a transfer area of 14 x 29 cm. The transfer time is 4 hours or overnight.
BIO-5000 Plus VIS gel scanner
The BIO-5000 Plus VIS gel scanner is a dual platform scanner specially designed to detect electrophoresis gels and blots up to 216 mm x 254 mm. It is equipped with energy-saving LEDs and an optical CCD sensor with a resolution of up to 4800 dpi. A variable optical density between 0.05 and 3.77 OD supports the recording of plane differences of electrophoresis gels.
GeneGnome XRQ
The GeneGnome XRQ is an imaging system for chemiluminescence. For this purpose, it is equipped with high performance and automation. For example, the highly efficient CCD camera allows an even more sensitive recording and the simple setup process automatically captures a high quality image of each Western blot. By built-in white light LEDs, colorimetric markers can be captured as well.
YouTube Videos about Western blotting
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