Bcl-2 ELISA Kit (B-Cell Lymphoma 2)

INTENDED USE

Human Bcl-2 ELISA kit is a procedure intended for measuring in vitro quantitative amounts of B-cell lymphoma 2 (Bcl-2) in human cell culture supernatant, plasma, serum, cell lysates, tissue homogenates and many other biological fluids. This assay has a minimum sensitivity limit of < 1.15 U/ml.

BACKGROUND

B-cell lymphoma 2 (Bcl-2) is vital in regulating apoptosis by either inhibiting (anti-apoptotic) or by inducing (pro-apoptotic). It is specifically referred to as crucial anti-apoptotic protein but not a proto-oncogene since it is unable to transduce growth signals. It has the ability to initiate a new gene family that is involved in the regulation of cell death and survival without affecting cell proliferation. This type of precise regulation and maintenance of balance between cell death and cell proliferation in multicellular organisms is very important for tissue homeostasis.

Bcl-2 gene is encoded by a proto-oncogene and it is described as an intracellular membrane associated protein which essentially functions to block programmed cell death. It has a unique function among many mammalian oncogenes since it is involved in the negative regulation of apoptosis, where a decrease in its levels can lead to cell death by apoptosis.

Overexpression of it is able to protect cells from death, however, this is neither able to lead to tumorgenic transformation of immortalized cells or immortalize normal cells. There have a many homologs which have been discovered, some of them are able to function as inhibitors of cell death whereas others can promote the process of apoptosis.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

• One 96-Well Plate: Pre-coated with anti Bcl-2 antibody.
• Standards: Concentrations 0, 1.562, 3.125, 6.25, 12.5, 25, 50, 100 ng/ml, made up from lyophilized recombinant.
• Sample/Standard Dilution Buffer.
• Biotinylated-Labelled Antibody.
• Antibody Dilution Buffer.
• HRP-Streptavidin Conjugate (SABC).
• SABC Dilution Buffer.
• TMB Substrate.
• Wash Buffer (25x).
• Plate Sealer.
• Product Instructions.

SENSITIVITY

The minimum detection sensitivity level of a B-cell lymphoma 2 (B-cell leukemia) using this Bcl-2 ELISA kit was 0.938 ng/ml. The dynamic range for this assay is 1.0 – 700.0 U/ml.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for Bcl-2, no cross reactivity or interference between Bcl-2 and analogues was detected.
– Recovery: Serum (85 – 101%), EDTA Plasma (86 – 105%), Heparin Plasma (87 – 103%).
– Linearity: Serum (92 – 104%), EDTA Plasma (82 – 99%), Heparin Plasma (83 – 97%).
– Precison Intra-Assay: CV < 8%.
– Precison Inter-Assay: CV < 10%.
– Stability: Less than 10%.

REFERENCES

1. Discoveries and controversies in BCL-2 protein-mediated apoptosis. FEBS J. (2016) 283 (14): 2690-700. Review. Zheng J.H., et al.
2. Targeting the B-cell lymphoma/leukemia 2 family in cancer. J Clin Oncol. (2012) 30 (25): 3127-35. Review. Davids M.S. and Letai A.
3. Cell death regulation by B-cell lymphoma protein. Apoptosis. (2006) 11 (4): 459-71. Review. Verma Y.K., et al.
4. Hematologic malignancies: newer strategies to counter the BCL-2 protein. J Cancer Res Clin Oncol. (2016) 142 (9): 2013-22. Review. Ebrahim A.S., et al.

ADDITIONAL INFORMATION

• Full Name: Bcl-2 ELISA Kit (B-Cell Lymphoma 2)
• Reactivity: Human
• Sample Type: Plasma, Biological Fluids, Serum, Tissue Homogenates
• Sensitivity: 0.938 ng/ml

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