Reduce The Background, Not The Specific Staining
Reliable time tested solution to unmask antigen on formalin-fixed sections
Any antibody preparation has some potential to produce non-specific reaction in the assay. This originates from:
- non-specific antibodies that are present in some proportion in any polyclonal antibody preparation, including affinity purified ones (often "affinity purified" means only isolation of IgG fraction on Prot A/G column, not the purification on the antigen column under very stringent conditions)
- low specificity antibodies among specific ones in polyclonal
- fragments of fallen apart IgGs in stored preparations, including monoclonal
- separate heavy and light chains of specific antibodies, produced by most hybridomas
All these are capable of binding non-specifically to molecules on tissue sections, blots, fixed cells and other objects for immune detection. In case of retrieved formalin sections the risk of non-specific reaction is even increased, since to active the epitope recovery the proteins comprising the tissue sections are denatured during HIER, thus making accessible many domains that are charged and are capable of binding the test immunoglobulins on non-specific manner.
The standard means to block non-specific binding of specific antibody preparation is to add some irrelevant protein, such as BSA, other serum, casein, etc. However, everyone who tried to do this knows that increasing (for effective blocking) concentration of such blocking agent leads to a great reduction of specific reaction as well. This is due to large blocking molecules binding to accessible sites on section and thus sterically blocking access of specific antibodies to epitopes of interest (schematically represented in the figure on the left, top). All our buffers developed for immune assays contain instead short (0.6-2 kD) peptides that are capable of block effectively non-specific reaction while not affecting the specific binding of antibody.
The presented collection of Immunohistology buffers has also some other benefits (see below) and allows you to achieve the best quality IHC result without compromising the antigen detection. The buffers can also be used in other immune assays, such as immunofluorescence on sections, flow cytometry on fixed cells, western blot, hybridization of sections with antibody detection.
The Retriever IHC buffers empower you to control non-specific staining on every step of immunohistochemistry. They are especially highly recommended for research pathology where, in contrast to diagnostics, many polyclonal and/or low-affinity antibodies are used.
All buffers available in 50 ml, 125 ml and 500 ml package. Ready to use.
A new class of blocking solutions based on chemically modified and fragmented ultra-pure casein. Effectively reduces unwanted binding of primary antibody and conjugates you use to charged surface of the slide and tissue section. Greatly reduces non-specific binding while preserving the specific reaction, by saturating potential non-specific protein-protein interactions. Moreover, in contrast to BSA-based, IgGm casein or serum - based blocking solutions there is no interaction of specific antibody and blocking protein itself. or oth Is not comparable to other commercially available or home-made blocking solutions. Recommended for research and diagnostic pathology, especially for retrieved sections and polyclonal antibodies.
Buffer for diluting your primary and secondary antibodies, especially if they were stored for a while, even at -20 in glycerol, or in refrigerator. Nonspecific binding of the antibodies, negative effects of disturbing substances and low or medium affinity cross-reactivities of the antibodies will be minimized, making your result more reliable. Excellent for IHC (frozen and formalin sections), flow cytometry on fixed cells, Western Blot and other immune assays.
When used in pathology, in also greatly reduces non-specific reactivity of human serum components and immunoglobulins in tissue, vessels and cells with mouse antibodies used on section.
For especially "trouble"-giving antibodies, as well as for in situ PCR applications, this dilutent may also be used as a washing buffer, preventing secondary binding of your analystes during washing.
Specifically designed for preparing solution of your HRP-conjugate used as the detection reagent. It is the Antibody-dilutent buffer with additional component for stabilizing your HRP-conjugate. Allows you to further standardize the assay preparing ready-to-use conjugate solutions in advance and store them in refrigerator without loss of activity.
|62710||Section Block||50 ml||150.00||Add to Cart|
|62711||Section Block||125 ml||265.00||Add to Cart|
|62712||Section Block||500 ml||995.00||Add to Cart|
|62713||Antibody Diluent||50 ml||150.00||Add to Cart|
|62714||Antibody Diluent||125 ml||325.00||Add to Cart|
|62715||Antibody Diluent||500 ml||1,250.00||Add to Cart|
|62713-01||Antibody Diluent for Frozen Sections||50 ml||152.00||Add to Cart|
|62714-01||Antibody Diluent for Frozen Sections||125 ml||325.00||Add to Cart|
|62715-01||Antibody Diluent for Frozen Sections||500 ml||1,250.00||Add to Cart|
|62716||HRP Conjugate Diluent||50 ml||102.00||Add to Cart|
|62717||HRP Conjugate Diluent||125 ml||320.00||Add to Cart|
|62718||HRP Conjugate Diluent||500 ml||1,150.00||Add to Cart|
|62718-15||Slide Washing Buffer||500 ml||130.00||Add to Cart|
Why the Universal Buffer?
Epitope recovery on formalin-fixed, paraffin embedded tissue sections, requires heat-induced treatment in buffer or, sometimes, proteolitic treatment of the deparaffinized tissue section. Which buffer to use, greatly depends on the exact antibody and the properties of the recognized epitope, therefore, one can find in literature and practice use of many buffers, including Citrate pH 3.4, Citrate 6.0, EDTA 8.0, Tris 9.0- 10.0, Tris-EDTA, etc.
Moreover, for individual antigens also the time of recovery in the individual buffer should be defined, as this may be different, and the treatment often destroys the epitope.
EMS offers a novel (patent-pending) technology for epitope recovery, primarily based on reversing the fixation effects of formaldehyde, which created links primarily between Îµ-amino groups of lysine and other amino-groups.
The buffer was extensively tested in pathology Departments in United Kingdom, and has shown excellent results when used with different antibodies, including those that normally require for successful staining treatment only in Low, or only High pH buffers, or require protolithic treatment.
The EMS Universal Buffer may be used in any heat-induced epitope recovery system (the time and temperature of treatment should be tested), but was specifically adjusted for tissue sections processing in our Antigen Retriever (62700 Series).
Using our Universal Buffer in our Retriever guarantees the highest rate of success in recovering epitope for any antibody, especially one that was never previously used on formalin-fixed sections.
Clear, non-toxic solution.
R-Universal Buffer is supplied as 10x concentrate. For epitope recovery dilute 1 part of stock with 9 parts of deionized water.
For epitope recovery dilute 1 part of stock with 9 parts of deionized water.
Stability and Storage
The preparation is stable for 1 year when stored unopened at +4°C. Every lot is issued with a certificate indicating the expiry date.
After opening, store at +4°C in the refrigerator and use within 6 months.
Each lot is certified for compliance to specifications. The product is produced under DIN EN ISO 9001 :2008 Quality Management system for the products in Immunoassay Development and Measurement, Products for Bioanalytics and lmmunoassays.
|62719-10||R-UNIVERSAL Epitope Recovery Buffer (10x stock)||125 ml||130.00||Add to Cart|
|62719-20||R-UNIVERSAL Epitope Recovery Buffer (10x stock)||500 ml||210.00||Add to Cart|