Shop All General Immunoassay Buffers & Reagents

ELISA Wash Buffer (30X) (Thermo Scientific™)

ELISA Wash Buffer is a Tris-based wash buffer for use in cytokine and other sandwich ELISA procedures. The buffer is the same formulation that is supplied with most Thermo Scientific Pierce ELISA Kits for cytokines and chemokines. The 30X concentrated stock solution is easily diluted 1:30 (vol:vol) in ultrapure water to yield sufficient wash buffer for a typical 96-well microplate ELISA protocol. Supplement an existing ELISA Kit or use as a convenient general purpose wash buffer for customized ELISA methods.

Features of the ELISA Wash Buffer:

Size: 50 mL (makes 1.5L of 1X buffer)
Storage: Shipped at on ice packs. Store at -20°C immediately upon receipt.
Application: Use as a wash buffer when performing ELISAs. This wash buffer is used in most, but not all, Thermo Scientific Pierce ELISA Kits. Please contact us for kit-specific information
Preparation: Thaw at room temperature (22-25°C). In a clean glass or plastic container, dilute the ELISA Wash Buffer concentrate 1:30 (vol:vol) with distilled or deionized water. Mix thoroughly. Equilibrate 1X wash buffer to room temperature before use in an assay. Prepare new 1X wash buffer for each day of use.
Automated Plate Washers: Storing the wash buffer in an automated system can result in contamination and poor ELISA performance. Automated plate washers must be cleaned regularly.

Guardian™ Peroxidase Conjugate Stabilizer/Diluent (Thermo Scientific™)

Thermo Scientific Guardian Peroxidase Conjugate Stabilizer/Diluent preserves the functional integrity and activity of horseradish peroxidase conjugated antibodies and other proteins at very dilute concentrations for long-term storage.

Features of Guardian Peroxidase Conjugate Stabilizer/Diluent:

Preserves HRP activity—no significant loss of HRP activity over a six-month period at room temperature (1:1000 dilution) or 12 months at 4°C
Convenient—store ready-to-use dilutions (1:1000 to 1:100,000) that maintain enzyme activity in the refrigerator—no aliquoting or freezing
Assay compatible—simply add your favorite blocking buffer to create the ideal diluent for your HRP-based ELISA system or store the HRP conjugate as a 1:1000 stock solution for Western blots and dilute in the final assay buffer
Saves money—less expensive than competitor formulations or ordering new HRP conjugates

With Guardian Peroxidase Conjugate Stabilizer/Diluent, typical 1 mg/mL antibody or streptavidin peroxidase conjugates can be diluted as much as 100,000-fold for storage at 4°C. Most ELISA and blotting applications for HRP conjugates require dilution of typical 1 mg/mL stocks by at least 1000-fold (to 1 µg/mL). Sensitive assay systems, such as with chemiluminescent substrates, often require dilution up to 100,000-fold (10 ng/mL). The Guardian Solution enables these 1X working concentrations of HRP conjugate to be prepared in advanced and stored at 4°C for 12 months or at room temperature for six months.For blotting applications, it is usually best to prepare a concentration for storage that can be diluted 10-fold in assay buffer (which may contain blocking components) for final use.

Typical colorimetric ELISA procedures requires that 10 mL of HRP-conjugated secondary antibody be diluted 1000- to 5000-fold from a 1 mg/mL stock solution. That necessitates pipetting 2 to 10 µL of stock into a tube with 10 mL of diluent. More sensitive assay systems, such as Western blots with Thermo Scientific SuperSignal West Pico, Dura or Femto Chemiluminescent Substrates for HRP, require a second round of dilution to achieve the needed working concentration of HRP conjugate. With Guardian Stabilizer-Diluent, HRP conjugates can be pre-diluted and stored at concentrations that are either ready to use or easily diluted 10- to 100-fold for final use. Preparing larger volumes of pre-diluted stocks ensures consistency and greater accuracy for assays performed repeatedly day after day.

Support Pack Plus (Invitrogen™)

Buffer pack for 10 plates (plates included)

Reactivity/Species
Human, Mouse, Rat

Reported Application
ELISA

eBioscience™ IHC Antigen Retrieval Solution - High pH (10X) (Invitrogen™)

The IHC Antigen Retrieval Solution- High pH (10X) is designed for use during the heat induced epitope retrieval (HIER) step prior to immunohistochemistry on formalin-fixed paraffin embedded tissue sections. The use of this buffer in combination with heat (often by microwave, water bath, or pressure cooker) has been shown to restore the antigenicity of proteins modified during the formalin fixation of tissue. This buffer is supplied as a 10X stock solution. Dilution of this buffer to 1X using reagent grade water results in a 10 mM Tris, 1 mM EDTA solution, pH 9.0.

Reported Application
Immunohistochemical Staining of Formalin-Fixed Paraffin Embedded Tissue Sections

CellVue Diluent C (For CellVue™ Cell Labeling Kits) (Invitrogen™)

Diluent C is the Buffer required for the dilution of the dyes in the CellVue™ Cell Labeling Kits. Please refer to the specific labeling kit for instructions of use.

Reported Application
Flow Cytometric Analysis, Microscopy

Amplex™ Red/UltraRed Stop Reagent (Invitrogen™)

Amplex® Red/UltraRed Stop Reagent is designed for use with the Amplex® Red and Amplex® UltraRed fluorogenic substrates and assay kits. Amplex® Red/UltraRed Stop Reagent provides convenience and control by allowing the fluorescence signal-generating reaction to be terminated at a user-determined time point. After addition of the stop reagent, the fluorescence signal remains stable for at least 3 hours.

See our complete line of Fluorescence Microplate assays.

Amplex® UltraRed and Amplex® Red assay kits are sensitive biomolecular assays based on hydrogen peroxide–generating enzyme systems linked to peroxidase–mediated oxidation of the fluorogenic Amplex® UltraRed or Amplex® Red substrates. Typically, detection reactions are performed in microplate wells and are initiated by adding the fluorogenic Amplex® UltraRed or Amplex® Red substrate, resulting in a continuous fluorescence increase that proceeds for 30 minutes or more. Ultimately, unknown analyte concentrations are determined by comparing fluorescence intensities measured at a certain time point during the reaction to parallel measurements at the same time point on standard samples of known concentration. Clearly, it is critical to ensure that the timing of the standard and unknown sample measurements is the same.

Amplex® Red/UltraRed Stop Reagent is designed for use with both the Amplex® Red and Amplex® UltraRed fluorogenic substrates and assay kits. It is designed to terminate reactions containing up to 0.1 units/mL of horseradish peroxidase (HRP) and 5 μM hyrogen peroxide. The fluorescent signal then remains stable for at least 3 hours.

Pierce™ Protein-Free T20 (PBS) Blocking Buffer (Thermo Scientific™)

Pierce Protein-Free Blocking Buffer is a non-protein compound that provides effective blocking for membrane-based and plate-based protein detection methods, resulting in extremely low background. It is supplied in a ready-to-use format.

Compare and view all available blocking buffers

Features of Protein-Free Blocking Buffer:
Protein-free blocker—minimizes or eliminates cross-reactivity associated with protein-based blocking buffers
Applications—western blotting (membranes), ELISA (microplates), and arrays (coated glass slides)
Streptavidin-friendly—free of biotin; no interference with avidin-biotin detection systems
High-performance—optimized and validated in many protein methods to provide high signal-to-noise ratio (i.e., no quenching of specific binding and signal but eliminating nonspecific binding and background)

Traditional blocking buffers contain proteins that can cross-react in certain immunodetection systems, resulting in high background and reduced signal. Pierce Protein-Free Blocking Buffers are devoid of protein while remaining highly effective at blocking plates, membranes, and other surfaces for ELISA, western blotting, glass slide arrays, and other applications.

Related products
Pierce Protein-Free (PBS) Blocking Buffer
Pierce Protein-Free (TBS) Blocking Buffer
Pierce Protein-Free T20 (TBS) Blocking Buffer

Pierce™ Clear Milk Blocking Buffer (10X) (Thermo Scientific™)

Pierce Clear Milk Blocking Buffer (10X) is a long-shelf life, high-performance replacement for homemade non-fat milk blocking buffers in western blotting applications.

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Our Clear Milk Blocking Buffer (10X) is a pre-formulated milk solution, clarified and stabilized for blocking excess nonspecific binding sites, reducing background in western blotting applications and diluting antibodies when used with nitrocellulose and PVDF membranes. It provides lower background, enhanced sensitivity, extended shelf life, and reproducible results compared to homemade buffers based on dissolved non-fat dry milk. Not recommended for avidin-based techniques because it contains some endogenous biotin.

StartingBlock™ (TBS) Blocking Buffer (Thermo Scientific™)

StartingBlock Blocking Buffer is a single purified protein for fast blocking of western blots and ELISA assays. It provides broad compatibility with a wide range of antibodies, antibody combinations, and other protein probing and assay systems. StartingBlock buffer is available in PBS and TBS with and without Tween20.

Compare and view all available blocking buffers

Features of StartingBlock Blocking Buffers:
Strip and reprobe without reblocking—blots stay blocked even after stripping with Thermo Scientific Restore Stripping Buffer (Cat. No. 21059)
Compatible with many detection systems—western blot, ELISA, and IHC with antibody or avidin/biotin probes (blocker is serum and biotin-free)
Short blocking times—less than 15 minutes for nitrocellulose or PVDF membranes, almost instantaneous for polystyrene microplate wells
Biotin- and serum protein-free formulation

Related products
StartingBlock (PBS) Blocking Buffer
StartingBlock T20 (PBS) Blocking Buffer
StartingBlock T20 (TBS) Blocking Buffer

Blocker™ Casein in TBS (Thermo Scientific™)

Blocker Casein is a ready-to-use solution of purified casein for blocking steps in western blot, ELISA, IHC, and nucleic acid detection methods. Blocker Casein is 1% (w/v) casein, which corresponds to the optimal concentration for most applications.

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Features:
Purified casein—single-protein blocking buffer provides fewer chances of cross-reaction with assay components than serum or milk solutions
Easy to use—1% casein solution is ready to use; can be diluted further as needed
Safe—stable, thimerosal-free formulation

Related products
Blocker Casein in PBS

InstantOne ELISA Cell Lysis Buffer (5X) (Invitrogen™)

Cell Lysis Buffer (5X) is included in the InstantOne ELISA kits. The solution is offered individually as some lab protocols require addition buffers.
Cell Lysis Buffer(5X) is used along with Enhancer Solution (Cat. No. IOES1) to make the Cell Lysis mixed used in the InstantOne phospho ELISA kits. The Cell Lysis Buffer (5X) contains a combination of detergents, phosphatase inhibitors, salts, and buffers. The Cell Lysis Buffer (5X) is supplemented with Enhancer Solution to yield a versatile Cell Lysis Mix that can be applied to many cells and tissues. Please note the difference in names. Cell Lysis Mix is referred to heavily in the InstantOne phospho ELISA assay protocol.
A. The Cell Lysis Mix (5X) is used to lyse cells in the presence of culture medium and is typically used to lyse non-adherent cells.
B. The 1X Cell Lysis Mix is prepared by simply diluting Cell Lysis Mix (5X) (a mixture of the Cell Lysis Buffer (5X) and the Enhancer Solution) 5-fold with water. This buffer is used to lyse cells after the removal of culture medium, and is typically used to lyse adherent cells or non-adherent cells that have been harvested by centrifugation. Lysis Mix should be used as the diluent for any dilution of cellular lysates that are required.
NOTE: *Supplementing Cell Lysis Mix with extra components (e.g. protease inhibitors, chelating agents, detergents) should be tested on a case-by-case basis for compatibility with InstantOne ELISA assays.

Reported Application
ELISA

Coating Buffer B

The Invitrogen Coating Buffer B is intended for use with Invitrogen Antibody Pairs as part of the recommended assay procedure.

eBioscience™ ELISA Wash Buffer, 1L packets (Invitrogen™)

The eBioscience ELISA Wash Buffer Powder is a dried powder formulation of phosphate buffered saline with 0.05% Tween-20. Each packet is reconstituted in 1 Liter of distilled, deionized water; the resulting 1x PBS/Tween-20 buffer (pH 7.4) is suitable for washing ELISA plate microwells. 10 packets of ELISA Wash Buffer Powder are supplied, yielding a total of 10 liters of 1x Wash Buffer.

Reported Application
ELISA

StartingBlock™ T20 (TBS) Blocking Buffer (Thermo Scientific™)

StartingBlock Blocking Buffer is a single purified protein for fast blocking of western blots and ELISA assays. It provides broad compatibility with a wide range of antibodies, antibody combinations, and other protein probing and assay systems. StartingBlock is available in PBS and TBS with and without Tween20.

Compare and view all available blocking buffers

Features of StartingBlock Blocking Buffers:
Strip and reprobe without reblocking—blots stay blocked even after stripping with Thermo Scientific Restore Stripping Buffer (Cat. No. 21059)
Compatible with many detection systems—western blot, ELISA, and IHC with antibody or avidin/biotin probes (blocker is serum and biotin-free)
Short blocking times—less than 15 minutes for nitrocellulose or PVDF membranes, almost instantaneous for polystyrene microplate wells
Biotin- and serum protein-free formulation

Related products
StartingBlock (PBS) Blocking Buffer
StartingBlock T20 (PBS) Blocking Buffer
StartingBlock (TBS) Blocking Buffer

Pierce™ Protein-Free (PBS) Blocking Buffer (Thermo Scientific™)

Pierce Protein-Free Blocking Buffer is a non-protein compound that provides effective blocking for membrane-based and plate-based protein detection methods, resulting in extremely low background. It is supplied in a ready-to-use format.

Compare and view all available blocking buffers

Features of Protein-Free Blocking Buffer:
Protein-free blocker—minimizes or eliminates cross-reactivity associated with protein-based blocking buffers
Applications—western blotting (membranes), ELISA (microplates), and arrays (coated glass slides)
Streptavidin-friendly—free of biotin; no interference with avidin-biotin detection systems
High-performance—optimized and validated in many protein methods to provide high signal-to-noise ratio (i.e., no quenching of specific binding and signal but eliminating nonspecific binding and background)

Traditional blocking buffers contain proteins that can cross-react in certain immunodetection systems, resulting in high background and reduced signal. Pierce Protein-Free Blocking Buffers are devoid of protein while remaining highly effective at blocking plates, membranes, and other surfaces for ELISA, western blotting, glass slide arrays, and other applications.

Related products
Pierce Protein-Free T20 (PBS) Blocking Buffer
Pierce Protein-Free (TBS) Blocking Buffer
Pierce Protein-Free T20 (TBS) Blocking Buffer