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Summary

One for All: the Universal Buffer

We proudly present patent pending R-Universal (R stands for Retriever) buffer for antigen unmasking/epitope recovery on formalin-fixed, paraffin embedded sections. Being specifically developed for the 2100 Retriever, it was thoroughly tested in pathology research labs in the UK, and now is available for all users of Retriever and other researchers who work with routine tissue material. Moreover, the quality of recovery greatly depends on the quality of fixation and processing, in particular – length of fixation, quality of washing from the fixative, and the temperature of paraffin used. When one performs large series of stainings on collections from archives, formed over some time, there is a risk of differences in efficacy of retrieval.

An excellent solution for epitope recovery has been achieved with the 2100 Retiever, and the new R-Universal buffer. The specially designed program of heating-cooling in Retriever was from the very beginning created for the best results in reshaping the denatured epitopes (allowing their proper refolding). But the new Retrieval buffer combines the removal of formaldehyde-formed protein cross-links with the removal of Ca++. And no toxic agent, such as Citraconic anhydride, is used.

As a result, you can perform recovery of practically all epitopes in one buffer.

Which allows you:

  • treat all sections in one buffer, so no buffer is wasted, when you have to treat several small series of sections in different buffers
  • use one buffer not only instead of different High, Low, etc buffers, but also to recover epitopes that normally require protease (Trypsin, Proteinase K) treatment
  • use practically any antibody previously not tested (or unsuccessfully tested) on formalin-fixed sections.
  • perform IF staining formalin-fixed on sections, as autofluorescence, even produced by excess of unwashed formaldehyde, will go away during the section processing
  • perform multicolour IF on sections, even when using antibodies that normally require epitope recovery in different buffers
  • get the strongest staining, superior to any other methods for epitope recovery. You will have more confidence that what you see as negative – is indeed negative, and not a result of poor epitope recovery

In other words, you may have all the conveniences of IHC on frozen sections, while having excellent tissue morphology of formalin-fixed.

6704On the image you see staining with three standard antibodies from Dako, to Cyclin D1, Lambda chain, and GFAP. Each of these require individual buffer (Low, High, or protease treatment) for a proper epitope recovery to avoid a non-specific staining produced by the respective mAb. Left panel shows staining on sections processed with recommended buffer, on the ladt panel – with R-universal buffer. Please not a much better discrimination between positive and negative cells (and stronger staining) for CyclinD1 and Lambda chain.

References

  1. T.Mellows, S.Litvinov, G.Thomas, A novel universal epitope recovery buffer for formalin-fixed sections: (in preparation)
  2. Leong, T.Y.-M., and Leong, A.S.-Y. (2007). How does antigen retrieval work? Adv Anat Pathol 14, 129–131.
  3. Fowler, C.B., Evers, D.L., O’Leary, T.J., and Mason, J.T. (2011). Antigen retrieval causes protein unfolding: evidence for a linear epitope model of recovered immunoreactivity. J. Histochem. Cytochem. 59, 366–381.
  4. Emoto, K., Yamashita, S., and Okada, Y. (2005). Mechanisms of heat-induced antigen retrieval: does pH or ionic strength of the solution play a role for refolding antigens? J. Histochem. Cytochem. 53, 1311–1321.
  5. Shi, S.-R., Shi, Y., and Taylor, C.R. (2011). Antigen retrieval immunohistochemistry: review and future prospects in research and diagnosis over two decades. J. Histochem. Cytochem. 59, 13–32.
  6. Syrbu, S.I., and Cohen, M.B. (2011). An enhanced antigen-retrieval protocol for immunohistochemical staining of formalin-fixed, paraffin-embedded tissues. Methods Mol. Biol. 717, 101–110.
  7. Siomin, Y.A., Simonov, V.V., and Poverenny, A.M. (1973). The reaction of formaldehyde with deoxynucleotides and DNA in the presence of amino acids and lysine-rich histone. Biochim. Biophys. Acta 331, 27–32.
  8. Namimatsu, S., Ghazizadeh, M., and Sugisaki, Y. (2005). Reversing the effects of formalin fixation with citraconic anhydride and heat: a universal antigen retrieval method. J. Histochem. Cytochem. 53, 3–11.

 

Product Size Price
1.5mm

Type: Puncher Tip

TAT06-1.5 1 ea
 
1.5mm (10 x 15 ; 150 wells)

Type: Mold Kit

TAM01-1.5 1 ea
 
1.5mm (9 x 10 ; 90 wells)

Type: Premade Recipient Block

TAB06-1.5 1 ea
 
1mm

Type: Puncher Tip

TAT06-1 1 ea
 
1mm (10 x 12 ; 120 wells)

Type: Premade Recipient Block

TAB06-1 1 ea
 
1mm (10 x 17 ; 170 wells)

Type: Mold Kit

TAM01-1 1 ea
 
2mm

Type: Puncher Tip

TAT06-2 1 ea
 
2mm (6 x 10 ; 60 wells)

Type: Premade Recipient Block

TAB06-2 1 ea
 
2mm (7x 10 ; 70 wells)

Type: Mold Kit

TAM01-2 1 ea
 
3mm

Type: Puncher Tip

TAT06-3 1 ea
 
3mm (5 x 6 ; 30 wells)

Type: Premade Recipient Block

TAB06-3 1 ea
 
3mm (5 x 8 ; 40 wells)

Type: Mold Kit

TAB01-3 1 ea
 
5mm

Type: Puncher Tip

TAT06-5 1 ea
 
5mm (3 x 5 ; 15 wells)

Type: Mold Kit

TAB-01-5 1 ea
 
5mm (4 x 5 ; 20 wells)

Type: Premade Recipient Block

TAB06-5 1 ea
 
Antibody Diluent For Frozen Sections ready-to-use

Type: IHC Buffers

AP0475-050 50 ml
 
Antibody Diluent For Frozen Sections ready-to-use

Type: IHC Buffers

AP0475-125 125 ml
 
Antibody Diluent For Frozen Sections ready-to-use

Type: IHC Buffers

AP0475-500 500 ml
 
Antibody Diluent ready-to-use

Type: IHC Buffers

AP0742-050 50 ml
 
Antibody Diluent ready-to-use

Type: IHC Buffers

AP0472-125 125 ml
 
Antibody Diluent ready-to-use

Type: IHC Buffers

AP0472-500 500 ml
 
Antibody Diluent W ready-to-use

Type: Western Blot Buffers

AP0501-050 50 ml
 
Antibody Diluent W ready-to-use

Type: Western Blot Buffers

AP0501-125 125 ml
 
Antibody Diluent W ready-to-use

Type: Western Blot Buffers

AP0501-500 500 ml
 
Base mold

Type: Mold Kit

TABM06 1 ea
 
Basket

Type: Accessories

R21008 1 each
 
Basket Lid for Chambers

Type: Accessories

R21009 1 each
 
Coating Buffer 7.4

Type: ELISA

AP0511-500 500 ml
 
Coating Buffer 9.6

Type: ELISA

AP0510-500 500 ml
 
Cord Set EU

Type: Accessories

R21005 1 each
 
Embedding cassette

Type: Mold Kit

TAC06 500/PK
 
Full set with 5 Tips and 5 sized blocks

Type: Tissue Microarrayer Set

TAT06 1 set
 
Green Silicone Sealing Gasket

Type: Accessories

R21007 1 each
 
Guide for 1mm recipient block

Type: Puncher Tip

TAG06 1 set
 
Peroxidase Conjugate Diluent ready-to-use

Type: Western Blot Buffers

AP0473-050W 50 ml
 
Peroxidase Conjugate Diluent ready-to-use

Type: Western Blot Buffers

AP0473-125W 125 ml
 
Peroxidase Conjugate Diluent ready-to-use

Type: Western Blot Buffers

AP0473-500W 500 ml
 
Peroxidase Conjugate Diluent ready-to-use

Type: ELISA

AP0473-050E 50 ml
 
Peroxidase Conjugate Diluent ready-to-use

Type: ELISA

AP0473-125E 125 ml
 
Peroxidase Conjugate Diluent ready-to-use

Type: ELISA

AP0473-500E 500 m
 
Peroxidase Conjugate Diluent ready-to-use

Type: IHC Buffers

AP0473-050 50 ml
 
Peroxidase Conjugate Diluent ready-to-use

Type: IHC Buffers

AP0473-125 125 ml
 
Peroxidase Conjugate Diluent ready-to-use

Type: IHC Buffers

AP0473-500 500 ml
 
Plate Block E

Type: ELISA

AP0513-500 500 ml
 
Plate Block, No Components of Animal Origin

Type: ELISA

AP0512-125 125 ml
 
Retriever Unit

Type: Retriever Unit

R2100-EU 1 each
 
Sample & Antibody Diluent E ready-to-use

Type: ELISA

AP0514-050 50 ml
 
Sample & Antibody Diluent E ready-to-use

Type: ELISA

AP0514-125 125 ml
 
Sample & Antibody Diluent E ready-to-use

Type: ELISA

AP0514-500 500 ml
 
Section Block ready-to-use

Type: IHC Buffers

AP0471-050 50 ml
 
Section Block ready-to-use

Type: IHC Buffers

AP0471-125 125 ml
 
Single set for 1 mm puncher with a guide

Type: Tissue Microarrayer Set

TAT06T1 1 set
 
Single set for 1.5mm puncher

Type: Tissue Microarrayer Set

TAT06T1.5 1 set
 
Single set for 2mm puncher

Type: Tissue Microarrayer Set

TAT06T2 1 set
 
Single set for 3mm puncher

Type: Tissue Microarrayer Set

TAT06T3 1 set
 
Single set for 5mm puncher

Type: Tissue Microarrayer Set

TAT06T5 1 set
 
Slide Chamber

Type: Accessories

R21001 Set of 3
 
Stripping Buffer ready-to-use W

Type: Western Blot Buffers

AP0503-050 50 ml
 
Stripping Buffer ready-to-use W

Type: Western Blot Buffers

AP0503-125 125 ml
 
Stripping Buffer ready-to-use W

Type: Western Blot Buffers

AP0503-500 500 ml
 
Universal Buffer

Type: Universal Buffers

AP0530-125 125 ml, 10x
 
Universal Buffer

Type: Universal Buffers

AP0530-500 500 ml, 10x
 
Washing Buffer E PBS-Based 10x

Type: ELISA

AP0515-500 500 ml
 
Washing Buffer W Tris-Based 10x

Type: Western Blot Buffers

AP0502-500 500 ml
 
Western Block, BSA-Free ready-to-use

Type: Western Blot Buffers

AP0500-125 125 ml
 
Western Block, BSA-Free ready-to-use

Type: Western Blot Buffers

AP0500-500 500 ml