BNIP3L ELISA Kit

INTENDED USE

Human BNIP3L ELISA kit can measure concentrations of BCL2/Adenovirus E1B 19 kDa protein-interacting protein 3-like (BNIP3L) present is plasma, serum, biological fluids or tissue homogenate samples.

INTRODUCTION

BNIP3L is a pro-apoptotic mitochondrial protein that mediates cell death in response to hypoxia and ischemia. It belongs to the Bcl-2 protein family, key regulators of the intrinsic apoptosis pathway. First discovered in 2000, it was recognized to have strong homology to BNIP3, another death-inducing Bcl-2 member. Whereas BNIP3 is activated by hypoxia, BNIP3L uniquely responds to ischemic stresses – making it an intriguing player in tissue damage following stroke or heart attack.

Research over the past two decades reveals BNIP3L orchestrates destruction of mitochondria and cell demise through distinct mechanisms. Under normal conditions, it resides in the cytoplasm in an inactive, monomeric form. However, ischemia causes it to homodimerize and translocate to the outer mitochondrial membrane. Here, its N-terminal domain inserts into the membrane while the C-terminus protrudes into the cytoplasm. Embedded BNIP3L then operates via two pathways – first, by physically opening the mitochondrial permeability transition pore leading to loss of membrane potential. Second, by interacting with the GTPase OPA1 to disrupt crucial mitochondrial inner membrane fusion events. Together these actions fragment the mitochondrial network, leading to release of pro-death factors like cytochrome C and activation of downstream cell death executors.

Intriguingly, the unique responsiveness of BNIP3L to ischemic injury places it at center stage following stroke or myocardial infarction. Research indicates that deleting it is protective following ischemia in neuronal and cardiac tissue, promoting idea that targeted BNIP3L inhibition could preserve cell viability. Promisingly, small molecule screens uncover several compounds able to disrupt BNIP3L-dimerization and downstream apoptotic signaling. As researchers continue elucidating molecular control of BNIP3L-activation, future therapies directed against BNIP3L may attenuate tissue damage in ischemia-reperfusion related pathologies.

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- BNIP3L.
Standards: 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.

TYPICAL RESULTS

For this BNIP3L ELISA kit, it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 0.156, 0.312, 0.625, 1.25, 2.5, 5, 20 ng/ml
Reactivity: Human
Sensitivity: 0.094 ng/ml
Range: 0.156 – 10 ng/ml
Principle: Sandwich, Double Antibody
Application: Research Use Only.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for BNIP3L, no cross reactivity or interference between BNIP3L and analogues was detected.
– Recovery: Serum (91 – 105%), EDTA Plasma (91 – 102%), Heparin Plasma (88 – 99%).
– Linearity: Serum (91 – 105%), EDTA Plasma (90 – 101%), Heparin Plasma (87 – 99%).
– Precison Intra-Assay: CV < 8%
– Precison Inter-Assay: CV < 10%

REFERENCES

BNIP3L-mediated mitophagy is required for mitochondrial remodeling during the differentiation of optic nerve oligodendrocytes. Autophagy. (2021) 17 (10) :3140-3159. Yazdankhah M., et al.
Mitophagy in tumorigenesis and metastasis. Cell Mol Life Sci. (2021) 78 (8): 3817-3851. Poole L.P. and Macleod K.F.
Dimerization of mitophagy receptor BNIP3L/NIX is essential for recruitment of autophagic machinery. Autophagy. (2021) 17 (5): 1232-1243. Marinković M., et al.
The emerging, multifaceted role of mitophagy in cancer and cancer therapeutics. Semin Cancer Biol. (2020) 66: 45-58. Bhutia S.K., et al.

ADDITIONAL INFORMATION

Full Name: BNIP3L ELISA Kit (BCL2/Adenovirus E1B 19 kDa Protein-Interacting Protein 3-Like)
Reactivity: Human
Sample Type: Plasma, Tissue Homogenates, Serum, Biological Fluids
Sensitivity: 0.094 ng/ml

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