抗PINK1抗体 | Anti-PINK1 Antibody

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	Biological Strategies Validation. Western Blot: PINK1 Antibody [BC100-494] - Analysis of PINK1 in HeLa whole cell lysate with and without treatment of 10 uM CCCP. Image courtesy of an anonymous customer review.
	Biological Strategies Validation. Immunocytochemistry/Immunofluorescence: PINK1 Antibody [BC100-494] - Immunocytochemistry of PINK1 antibody (BC100-494 Lot G). HeLa cells were treated with valinomycin (1 uM for 24h) prior to being fixed in 10% buffered formalin for 10 min and permeabilized in 0.1% Triton X-100 in PBS for 10 min. Cells were incubated with BC100-494 at 20 ug/ml for 1h at room temperature, washed 3x in PBS and incubated with Alexa-Fluor488 anti-rabbit secondary antibody. PINK1 (Green) was detected at the mitochondria. Tubulin (Red) was detected using an anti-tubulin antibody with an anti-mouse DyLight 550 secondary antibody. DNA (Blue) was counterstained with DAPI. Note: mitochondria staining might not be easily observed without treatment with valinomycin or CCCP.
	Biological Strategies Validation. Immunocytochemistry/Immunofluorescence: PINK1 Antibody [BC100-494] - HeLa cells were treated with valinomycin (1 uM for 24h) prior to being fixed in 10% buffered formalin for 10 min and permeabilized in 0.1% triton X-100 in PBS for 10 min. Cells were incubated with BC100-494 at 20 ug/ml for 1h at room temperature, washed 3x in PBS and incubated with Alexa-Fluor488 anti-rabbit secondary antibody. PINK1 (Green) was detected at the mitochondria. Tubulin (Red) was detected using NB100-690 with an anti-mouse DyLight 550 secondary antibody. DNA (Blue) was counterstained with DAPI. Note: mitochondrial staining may only be visible after treatment with valinomycin or CCCP.
	Western Blot: PINK1 Antibody [BC100-494] - Analysis of PINK1 in mouse liver and hypatocytes using PINK1 antibody. Image from verified customer review.
	Genetic Strategies Validation. Western Blot: PINK1 Antibody [BC100-494] - WB analysis of lysates derived from hTERT-RPE1 cells transfected with non-targeting (NT1) or PINK1 targeting siRNA for 72 h. 20 uG were loaded. Image from verified customer review.
	Immunohistochemistry-Paraffin: PINK1 Antibody [BC100-494] - Rabbit heart tissue. Image from verified customer review.
	Western Blot: PINK1 Antibody [BC100-494] - Western blot image of PINK1 antibody (BC100-494) in multiple cells lines. Human HeLa (lane 1), Mouse NIH-3T3 (lane 2), L929 (lane 3) and Rat PC12 (lane 4) whole cell protein were separated by SDS-PAGE on a 7.5% polyacrylamide gel. Protein was transferred to PVDF membrane and probed with 2 ug/ml BC100-494 in 1% BSA and detected with an HRP-conjugated anti-rabbit secondary antibody using chemiluminescence. PINK1 was detected at approximately 60 kDa (arrowhead).
	Biological Strategies Validation. Western Blot: PINK1 Antibody [BC100-494] - Whole cell protein from HeLa cells treated without or with valinomycin (1 uM for 24h) as indicated were separated by SDS-PAGE on a 7.5% polyacrylamide gel. Protein was transferred to PVDF membrane and probed with 1.0 ug/ml BC100-494 in 1% BSA and detected with an HRP-conjugated anti-rabbit secondary antibody using chemiluminescence. PINK1 was detected at approximately 60 kDa in the treated sample(arrowhead).

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Background

PINK1 (PTEN induced putative kinase 1) is a mitochondrial serine/threonine kinase which maintains mitochondrial function/integrity, provides protection against mitochondrial dysfunction during cellular stress by phosphorylating mitochondrial proteins, and is involved in the clearance of damaged mitochondria via selective autophagy (mitophagy). PINK1 is synthesized as a 63 kDa protein which undergoes proteolytic processing to generate at least two cleaved forms (55 kDa and 48 kDa). PINK1 and its cleavage products have been found in the cytosol as well as in different sub-mitochondrial compartments, and according to reports; PINK1 may be targeted to OMM (outer mitochondrial membrane) with its kinase domain facing the cytosol, providing a possible explanation for the observed physical interaction with the cytosolic E3 ubiquitin ligase Parkin. Defective PINK1 may cause alterations in processing, stability, localization and activity as well as binding to substrates/interaction-partners which ultimately leads to differential effects on mitochondrial function and morphology. Mutations in PINK1 are linked to autosomal recessive early onset Parkinson's disease, and are associated with loss of protective function, mitochondrial dysfunction, aggregation of alpha-synuclein, as well as proteasome dysfunction.

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