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                SiteClick™ Antibody Azido Modification Kit Invitrogen™

                Create a label-ready site-specific azido-modified antibody without lengthy and inefficient genetic modification using the SiteClick Antibody Azido Modification Kit. SiteClick labeling uses enzymes to specifically attach an azido moiety to the heavy chains of an IgG antibody, ensuring that the antigen binding domains remain unaltered for binding to the antigen target. This site selectivity is achieved by targeting the carbohydrate domains present on essentially all IgG antibodies regardless of isotype and host species. Once azido–modified, a variety of labeled SiteClick sDIBO alkynes are available for attachment to the antibody via Click chemistry at your choice of scale. This provides the flexibility to choose different labels for your antibody depending on your assay.

                Features of the SiteClick Antibody Azido Modification Kit:
                • Contains everything required to azido-modify any species or isotype of IgG antibody
                • Easy-to-follow step-by-step protocol
                • Highly efficient, site-specific, reproducible labeling chemistry results in high quality antibody conjugate

                Learn more about SiteClick labeling technology ›

                Custom SiteClick Antibody Labeling Service and sDIBO labels
                If you have an antibody that is considered 'difficult to label' or has lost activity after labeling using a conventional method, please contact our custom service representatives to determine whether the SiteClick Antibody Labeling Service would be right for your antibody. We offer complete custom SiteClick antibody labeling services with the option of multiple detection molecules including biotin, Alexa Fluor dyes, Qdot fluorophores, R-PE, chelates for PET imaging, and many others.

                Western Devices Benchtop Bundle Invitrogen™

                The Western Devices Benchtop Bundle contains all the necessary devices, consumables, and reagents to perform high quality westerns.

                Contents of the Western Devices Benchtop Bundle:
                Mini Gel Tank
                Bolt 4-12% Bis-Tris Plus gels, 10-well
                Bolt MES Running Buffer (20X)
                Bolt LDS Sample Buffer (4X)
                Bolt Sample Reducing Agent (10X)
                SeeBlue Plus2 Pre-Stained Standard
                iBlot 2 Gel Transfer Device
                iBlot 2 Transfer Stacks, nitrocellulose, regular size
                iBind Western Device
                iBind Cards
                iBind Solution Kit
                SuperSignal West Pico Plus Chemiluminescent Substrate

                Mini Gel Tank
                The Mini Gel Tank is compatible with Novex gels, NuPAGE gels, and Bolt Bis-Tris Plus gels. Each Mini Gel Tank can accommodate up to two gels per run. The unique tank design enables convenient side-by-side gel loading and enhanced viewing during use. Optimized conditions using constant voltage allow for Bolt Bis-Tris Plus gels to be run in approximately 35 minutes.

                Bolt Bis-Tris Plus gels
                Bolt Bis-Tris Plus gels are precast polyacrylamide gels designed for optimal separation of a wide range of protein sizes under denaturing conditions. Bolt Bis-Tris Plus gels are designed to deliver consistent gel performance and provide a neutral pH environment that minimizes protein modifications. Bolt gels are ideal for western blot transfer and analysis, and any other techniques where protein integrity is crucial. Also use Bolt gels to obtain optimal results for your day-to-day protein separation needs.

                iBlot 2 Gel Transfer Device
                The iBlot 2 Gel Transfer Device performs western blotting transfer simply, efficiently, and reliably, within seven minutes without the need for liquid buffers. The iBlot 2 system is compatible with both polyvinylidene difluoride (PVDF) and nitrocellulose membranes, and has comparable performance to traditional wet transfer methods in a fraction of the time. The iBlot 2 Gel Transfer Device is an integral part of the iBlot 2 Dry Blotting system, which consists of the transfer device and consumable transfer stacks that contain the required buffers and transfer membrane.

                iBind Western Device
                The iBind Western Device is an automated western-processing device that performs every step in the western protocol from blocking to washes to antibody incubations via sequential lateral flow (SLF). SLF allows the timely release and flow of solutions and antibodies to the membrane without need of an external power source. Each solution is released from the iBind device wells to an iBind Card via SLF; the solutions are then wicked towards the stack region of the card. The glass fiber matrix of the card allows for homogenous and consistent flow of the solutions to the membrane, increasing the antigen-antibody interaction. The iBind western detection procedure minimizes hands-on time and is highly reproducible and compatible with all western protocols. Set up time is fast: in less than 3 hours you are ready for the detection step.

                SiteClick™ Biotin sDIBO Alkyne Invitrogen™

                SiteClick labeled sDIBO alkynes for antibody labeling are optimized for easy attachment to azido modified antibodies using copper-free Click chemistry. This labeled sDIBO alkyne can be used with antibodies that have been modified using the SiteClick Antibody Azido Modification Kit or engineered to contain azido moieties. These labeled sDIBO alkynes are improved versions of our original DIBO cyclooctynes, yielding conjugates that are less 'sticky' and that produce lower signal background in biological samples. This modular labeling system gives you the option to choose different fluorescent labels for your antibody and attach another molecule via streptavidin or your own molecule via amine-reactive or amine-containing moieties depending on your assay.

                Learn more about SiteClick labeling technology ›

                Custom SiteClick Antibody Labeling Service and sDIBO labels
                If you have an antibody that is considered 'difficult to label' or has lost activity after labeling using a conventional method, please contact our custom service representatives to determine whether the SiteClick Antibody Labeling Service would be right for your antibody. We offer complete custom SiteClick antibody labeling services with the option of multiple detection molecules including biotin, Alexa Fluor dyes, Qdot fluorophores, R-PE, chelates for PET imaging, and many others.

                PVDF Transfer Membrane, 0.45 µm, 26.5 cm x 3.75 m Thermo Scientific™

                Pierce PVDF Transfer Membrane, 0.45 µm, is made of high-quality polyvinylidene difluoride and provides high binding capacity for proteins and nucleic acids for western, southern, and northern blotting methods. The 0.45-µm pore size makes this membrane ideal for transfer for proteins >10 kDa.

                See all membranes ›

                Features:
                Membrane: polyvinylidene difluoride (PVDF)
                Binding capacity: goat IgG: 294 µg/cm2; BSA: 215 µg/cm2; insulin: 160 µg/cm2
                Re-probe characteristics: yes
                Pre-activation: required with 100% alcohol (methanol)
                Compatibility: compatible with commonly used transfer conditions and detection methods such as staining, chemiluminescence, and radiolabeling
                Durability: compatible with most organic solvents, acids, and mild bases

                SiteClick™ pHrodo™ iFL Red sDIBO Alkyne Invitrogen™

                SiteClick labeled sDIBO alkynes for antibody labeling are optimized for easy attachment to azido modified antibodies using copper-free Click chemistry. This labeled sDIBO alkyne can be used with antibodies that have been modified using the SiteClick Antibody Azido Modification Kit or engineered to contain azido moieties. These labeled sDIBO alkynes are improved versions of our original DIBO cyclooctynes, yielding conjugates that are less 'sticky' and that produce lower signal background in biological samples. This modular labeling system gives you the option to choose different fluorescent labels for your antibody and attach another molecule via streptavidin or your own molecule via amine-reactive or amine-containing moieties depending on your assay.

                The increase in fluorescence of pHrodo iFL red dye as the pH changes from basic to acidic correlates with the acidification of intracellular vesicles, making it a valuable tool for the study of antibody internalization via endocytosis or phagocytosis and regulation by environmental factors, drugs, or pathogens.

                Learn more about SiteClick labeling technology ›
                Learn more about IFL pHrodo antibody internalization ›

                Custom SiteClick Antibody Labeling Service and sDIBO labels
                If you have an antibody that is considered 'difficult to label' or has lost activity after labeling using a conventional method, please contact our custom service representatives to determine whether the SiteClick Antibody Labeling Service would be right for your antibody. We offer complete custom SiteClick antibody labeling services with the option of multiple detection molecules including biotin, Alexa Fluor dyes, Qdot fluorophores, R-PE, chelates for PET imaging, and many others.

                eBioscience™ Avidin HRP Invitrogen™

                The avidin horseradish peroxidase is commonly used as a second step for detection of biotinylated antibodies by ELISA and western blot. Avidin binds to biotin with high affinity. [Note: Sodium azide is an inhibitor of horseradish peroxidase enzymatic activity and should not be added to the AV-HRP diluent.]

                Conjugate
                HRP

                Reported Application
                Western Blotting, ELISA

                PVDF Transfer Membrane, 0.2 µm, 26.5 cm x 3.75 m Thermo Scientific™

                Pierce PVDF Transfer Membrane, 0.2 µm, is made of high-quality polyvinylidene difluoride and provides high binding capacity for proteins and nucleic acids for western, southern, and northern blotting methods. The 0.2-µm pore size makes this membranes ideal for protein analysis of small amounts of proteins (as little as 10 pmoles), amino acid analysis, and transfer of small molecular weight proteins and peptides.

                See all membranes ›

                Features:
                Membrane: polyvinylidene difluoride (PVDF)
                Binding capacity: goat IgG: 448 µg/cm2; BSA: 340 µg/cm2; insulin: 262 µg/cm2
                Re-probe characteristics: yes
                Pre-activation: required with 100% alcohol (methanol)
                Compatibility: compatible with commonly used transfer conditions and detection methods such as staining, chemiluminescence, and radiolabeling
                Durability: compatible with most organic solvents, acids, and mild bases

                ProLong™ Live Antifade Reagent, for live cell imaging Invitrogen™

                ProLong® Live Antifade Reagent helps prevent the loss of fluorescent signal due to photobleaching during live cell imaging. When dyes are illuminated, they can degrade (usually permanently), reducing the sample’s fluorescent intensity and leading to the creation of free radical singlet oxygen, an element that can further degrade neighboring dye molecules. ProLong® Live Antifade Reagent metabolizes this element, increasing the signal intensity and duration while keeping the background signal low. ProLong® Live reagent has little-to-no effect on cellular viability, proliferation, or other cellular functions in experiments up to 48 hours.

                Features of ProLong® Live Antifade Reagent include:
                • Photobleach protection in live cell experiments, for fluorophores such as GFP, RFP, Hoechst 33342, MitoTracker®, LysoTracker®, and CellTracker®dyes
                • Little-to-no effect on cellular viability, proliferation, or other cellular functions
                • Easy to use, just add to the imaging solution or complete cell culture media or buffer, incubate, and image

                ProLong® Live Antifade Reagent contains enzymes from the plasma membrane of naturally occurring E. coli. These enzymes metabolize the elements that can cause photobleaching. They work in the extracellular space and are not known to enter cells, so intracellular functions are minimally affected by ProLong® Live Antifade Reagent. As shown in figure below, cells grown in the presence of ProLong® Live Antifade Reagent display minimal effects on cellular viability or proliferation. ProLong® Live Antifade Reagent provides photobleach protection for most fluorophores frequently used in live cell fluorescent imaging, such as GFP, RFP, Hoechst 33342, MitoTracker®, LysoTracker® and CellTracker® dyes. ProLong® Live Antifade Reagent can be added to any cell culture media or buffer that is suitable for fluorescent imaging.

                SiteClick™ Alexa Fluor™ 488 sDIBO Alkyne Invitrogen™

                SiteClick labeled sDIBO alkynes for antibody labeling are optimized for easy attachment to azido modified antibodies using copper-free Click chemistry. This labeled sDIBO alkyne can be used with antibodies that have been modified using the SiteClick Antibody Azido Modification Kit or engineered to contain azido moieties. These labeled sDIBO alkynes are improved versions of our original DIBO cyclooctynes, yielding conjugates that are less 'sticky' and that produce lower signal background in biological samples. This modular labeling system gives you the option to choose different fluorescent labels for your antibody and attach another molecule via streptavidin or your own molecule via amine-reactive or amine-containing moieties depending on your assay.

                Learn more about SiteClick labeling technology ›

                Custom SiteClick Antibody Labeling Service and sDIBO labels
                If you have an antibody that is considered 'difficult to label' or has lost activity after labeling using a conventional method, please contact our custom service representatives to determine whether the SiteClick Antibody Labeling Service would be right for your antibody. We offer complete custom SiteClick antibody labeling services with the option of multiple detection molecules including biotin, Alexa Fluor dyes, Qdot fluorophores, R-PE, chelates for PET imaging, and many others.

                Owl™ VEP-3 Large Tank Electroblotting System Thermo Scientific™

                Rapidly transfer high molecular weight proteins with the Thermo Scientific? Owl? VEP-3 Large Tank Electroblotting System.

                Low-Fluorescence PVDF Transfer Membranes, 0.2 μm,m, 7 x 8.4 cm Thermo Scientific™

                Pierce Low-Fluorescence PVDF Transfer Membranes are made of high-quality polyvinylidene difluoride and provide high binding capacity for proteins and nucleic acids for western, southern, and northern blotting methods. This low-fluorescence PVDF membrane has a 0.2-micron pore size and is ideal for use with fluorescent probes.

                See all membranes ›

                Features of low-fluorescence PVDF membranes:
                Membrane: polyvinylidene difluoride (PVDF)
                Re-probe characteristics: yes
                Pre-activation: required with 100% alcohol (methanol, ethanol, or isopropanol)
                Compatibility: compatible with commonly used transfer conditions and detection methods such as staining, immunodetection, chemiluminescence, fluorescence, and radiolabeling
                Durability: compatible with most organic solvents, acids, and mild bases
                Binding interactions: hydrophobic

                iBind™ Flex Midi Insert Replacement Invitrogen™

                The iBind™ Flex Midi Insert Replacement is an accessory for the iBind™ Flex Western Device to process one midi blot.

                iBind Flex Western System
                The iBind™ Flex Western System offers automated western blot processing and can accommodate various blot formats including midi, mini, and vertically cut strips. Based on Sequential Lateral Flow (SLF) technology, all blocking, antibody incubation and washing steps during immunodetection are hands-free. Compared to manual methods, the iBind Flex Western System requires less primary antibody, enables reproducible results and is compatible with downstream detection protocols.

                XCell SureLock™ Mini-Cell and XCell II™ Blot Module Invitrogen™

                The XCell SureLock Mini-Cell and XCell II Blot Module set is an electrophoresis and blotting system for mini gels. It includes the XCell SureLock Mini-Cell for running Invitrogen mini gels and the XCell II Blot Module for blotting up to two mini gels at a time.

                See all available electrophoresis chamber systems ›

                How the system works
                The XCell SureLock Mini-Cell holds gels firmly with a simple gel tension wedge. When the lever on the gel tension wedge is pushed forward into the locked position, an even, horizontal force is generated. This seals the gel/buffer core assembly firmly into position in the lower buffer chamber. The positive locking action of the gel tension wedge ensures a trouble-free, leak-free gel run every time.

                XCell II Blot Module
                The XCell II Blot Module allows you to easily transfer proteins or nucleic acids from mini gels to membranes. It fits into the XCell SureLock and XCell II Mini-Cells in place of the gel/buffer core assembly. It requires less than 200 mL of transfer buffer for western, southern, and northern transfers. Tough platinized titanium and stainless steel electrodes create a uniform electrical field without clamps or hinged gel holders. Maximum blot size is 9 cm x 9 cm.

                Upgrade your XCell II to an XCell SureLock Mini-Cell
                Conversion of an XCell II into an XCell SureLock is easy with the SureLock Retrofit Kit (available separately). The kit consists of a SureLock mini-cell lid, gel tension wedge, and molded mini-gel buffer dam.

                Pierce™ Background Eliminator Kit Thermo Scientific™

                The Thermo Scientific Pierce Background Eliminator Kit provides fast, easy removal of haze and artifacts on developed X-ray film to correct for overexposure of blots, whether from chemiluminescent Western blot or DNA or RNA autoradiography.

                Features of the Background Eliminator Kit:

                For overexposed X-ray film—reverses development of silver-based autoradiography film removing silver emulsion from the film surface
                Recovers and preserved data—evenly reduces developed signal over the entire film so relative intensity (densitometry) of bands is not altered
                Optimizes for signal-to-noise—allows readjustment of overdeveloped film to obtain the best image with respect to background haze, speckles and oversaturated bands that need to be resolved
                Saves time and resources—eliminates the need to repeat experiments or re-expose a new piece of film to obtain an optimal image

                This two-reagent kit is simple to use as demonstrated in the video below. Mix the two reagents (1:1 ratio) and immerse a developed piece of film (recent or old) in the working solution. Watch for 1 to 5 minutes until the desired image appears, then stop the reaction by rinsing the film in water. Pierce Background Eliminator for Film evenly reverses development-signal over the entire film, preserving relative densitometry values so that analysis and publication of results is still valid and possible.

                Applications:
                • X-ray film developed from enhanced chemiluminescence Western blots of any kind, whether from Thermo Scientific SuperSignal, Pierce ECL or other commercial substrate
                • X-ray film developed from any chemiluminescent or radioactive gel-shift assays, including Thermo Scientific LightShift Chemiluminescent EMSA Kits for DNA or RNA
                • Autoradiography film developed from chemiluminescent or isotopic (32P or 35S) Northern or Southern blots, including with Thermo Scientific North2South Chemiluminescent Detection Kit
                • Any developed silver-emulsion photographic film, whether blue, clear, single- or double-sided, and regardless of the method of exposure

                High background, shading, overexposed bands and speckling are problems inherent to film exposure. High background and shading are caused by overexposure, poor use of blocking buffer or inappropriate enzyme-labeled probe or antibody concentration. Overexposed bands are a common occurrence when the enzyme-labeled probe or antibody used is too concentrated or if the film was exposed for too long. Speckling and shading occur when enzyme conjugates form complexes and precipitate on the blot. Pierce Background Eliminator can correct all these problems without the need to re-expose a blot to film or re-do an entire experiment. The working solution can be used with newly exposed films or those that have been stored for years. In addition, the product can be used with any brand of film.

                SuperSignal™ West Pico PLUS Chemiluminescent Substrate Thermo Scientific™

                Thermo Scientific SuperSignal West Pico PLUS Chemiluminescent Substrate is an enhanced chemiluminescent (ECL) horseradish peroxidase (HRP) substrate that enables picogram to high femtogram-level protein detection by western blot analysis.

                SuperSignal West Pico PLUS Chemiluminescent Substrate characteristics include:
                Sensitivity: pictogram to femtogram sensitivity
                Stability: 8-hr working solution stability; 1-year kit stability at room temperature
                Compatibility: nitrocellulose and PVDF membranes
                Signal duration: 6 to 24 hours
                Recommended primary antibody concentration: 1:1,000–1:5,000 dilution (0.2–1.0 µg/mL)
                Recommended secondary antibody concentration: 1:20,000–100,000 dilution (10–50 ng/mL)

                Compare and learn more about our western blot chemiluminescent substrates ›
                View all HRP substrates available ›

                SuperSignal West Pico PLUS substrate is designed to provide excellent signal intensity and sensitivity for western blotting with HRP conjugates. The intensity of the light emission combined with the exceptional duration allows for the acquisition of multiple exposures to more easily obtain publication-quality blot images. SuperSignal West Pico PLUS substrate is compatible with different membranes, blocking reagents, and a wide range of antibody dilutions, making it an ideal choice for most western blotting applications.

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