First slide
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Product summary

  • Technology FISH
  • Application Solid tumour
  • Areas of interest Lymphoma
  • Region 18q21.33-q22.1
  • Label    
  • Product Code LPS 028 (10 tests)
    LPS 028-S (5 tests)
  • Regulatory Status In vitro diagnostic. This product is intended to be used on Carnoy’s solution (3:1 methanol/acetic acid) fixed haematological samples, or formalin-fixed paraffin-embedded (FFPE) tissues. Disease information supported by the literature and is not a reflection of the intended purpose of this product.

Chromomaps

Overview

Probe specification

  • BCL2, 18q21.33-q22.1, Red
  • BCL2, 18q21.33-q22.1, Green

The BCL2 product consists of two green 161kb, 196kb probes and two red 304kb, 176kb probes, which are positioned on each side of the BCL2 gene.

 

Probe information

The B-cell CLL/Lymphoma 2 (BCL2) gene located at 18q21.33 encodes one of a large protein family that regulates and contributes to programmed cell death, or apoptosis, by controlling mitochondrial membrane permeability1.

Translocations of the BCL2 gene result in constant expression of the BCL2 protein. BCL2 usually translocates to the immunoglobulin (IG) heavy chain (IGH) gene via a t(14;18)(q32;q21.3) and rarely to IG light chain (IGK or IGL) loci via t(2;18)(p11;q21.3) or t(18;22)(q21.3;q11)2.

The translocation (t(14;18)(q32.33;q21.33)) is thought to be brought about by an error in the joining function of the IGH gene, mediated by the observation that both IGH and BCL2 are arranged next to each other in 3D space in normal B lymphocytes3. The translocation breakpoint at the end of the Joining (J) segment, and the subsequent fusion of the BCL2 gene to this region, results in the BCL2 gene coming under the regulatory control of those processes normally involved in maintenance of IGH gene activity4.

The t(14;18) is observed in 70-95% of Follicular Lymphoma (FL) cases and 20-30% of Diffuse Large B-cell Lymphoma (DLBCL)2. Presence of the t(14;18) translocation in DLBCL is a predictor of outcome and has a poor prognostic effect5. BCL2 translocations have also been implicated in Chronic B-cell Lymphoproliverative Disease (CLPD) and occur frequently in Chronic Lymphocytic Leukaemia (CLL)6.

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References

  1. Sharpe et al., Biochim Biophys Acta. 2004;1644(2-3):107-13
  2. Tomita N., J Clin Exp Hematop 2011;51(1):7-12
  3. Roix et al., Nat Genet 2003;34(3):287-91
  4. Stoos-Veić et al., Coll Antropol. 2010 Jun;34(2):425-9
  5. Barrans et al., Clin Cancer Res 2003; 9; 2133
  6. Bassegio L et al., Br J Haematol 2012;158(4):489-98

FFPE tissue preparation and FISH protocol

Select a protocol step to view:

Heat pretreatment

Icon representing the heat pre-treatment stage of the fluorescence in situ hybridisation (FISH) protocol for FFPE samples.
  • Heat 50ml Tissue Pretreatment Solution (Reagent 1) in a porcelain wash jar or coplin jar immersed in a waterbath until it is either boiling or 98 - 100°C.
  • Boil slides for 30 minutes (Note: different incubation times may be required depending on tissue fixation. A 30-minute incubation is a recommended starting point).
  • Wash in PBS or dH2O at room temperature (RT) for 2 x 3 minutes.
Solid tumour FFPE FISH protocol Video Image
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Solid tumour FFPE FISH protocol

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