Skip to Content
MilliporeSigma
  • Protocol for labeling and fixation of intact lysosomes with esterified amino acid analogs to assess lysosomal expansion in living eukaryotic cells.

Protocol for labeling and fixation of intact lysosomes with esterified amino acid analogs to assess lysosomal expansion in living eukaryotic cells.

STAR protocols (2021-11-11)
Gianluca Scerra, Maria Gabriella Caporaso, Maurizio Renna, Massimo D'Agostino
ABSTRACT

The lysosomal compartment is a key hub for cell metabolism, and morphological alterations have been described in several pathological conditions. Here, we describe the use of amino acid analogs modified by the presence of a methyl ester group that accumulates within lysosomes. This generates an intraluminal osmotic effect able to trigger a rapid and selective expansion of the lysosomal compartment within 2 h of treatment. We also present protocols to preserve lysosomal morphology, which yields a more accurate size measurement. For complete details on the use and execution of this protocol, please refer to Scerra et al. (2021).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
L-Alanine methyl ester hydrochloride, 99%
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
25 g
In Stock
Details...
$40.20
100 g
In Stock
Details...
$81.20
Sigma-Aldrich
L-Leucine methyl ester hydrochloride, 98%
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
25 g
In Stock
Details...
$40.20
100 g
In Stock
Details...
$81.20
Sigma-Aldrich
Saponin, for molecular biology, used as non-ionic surfactant
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
25 g
In Stock
Details...
$40.20
100 g
In Stock
Details...
$81.20
Sigma-Aldrich
Bovine Serum Albumin, cold ethanol fraction, pH 5.2, ≥96%
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
25 g
In Stock
Details...
$40.20
100 g
In Stock
Details...
$81.20
Sigma-Aldrich
Methanol, suitable for HPLC, ≥99.9%
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
25 g
In Stock
Details...
$40.20
100 g
In Stock
Details...
$81.20
Sigma-Aldrich
HEPES, ≥99.5% (titration)
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
25 g
In Stock
Details...
$40.20
100 g
In Stock
Details...
$81.20
Sigma-Aldrich
Glutaraldehyde solution, Grade I, 70% in H2O, specially purified for use as an electron microscopy fixative or other sophisticated use
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
25 g
In Stock
Details...
$40.20
100 g
In Stock
Details...
$81.20
Sigma-Aldrich
L-Histidine methyl ester dihydrochloride, 97%
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
25 g
In Stock
Details...
$40.20
100 g
In Stock
Details...
$81.20