Neuro2a Chloroquine Treated / Untreated Cell Lysate Summary
NBP2-49688 contains 1 vial each of 0.1ml of Neuro2a Chloroquine treated and 0.1ml of Neuro2a untreated cell lysate. This lysate set is useful as a positive and negative controls for targets such as LC3, p62 etc. The researchers should use anti-mouse primary antibodies when working with these lysates.
Autophagy/LC3 Positive Control
Neuro2A cells were cultured under standard laboratory condition until semi-confluent (70-80%). The cells were then treated with or without Chloroquine to 50 uM for 24 hours. The cells were washed in PBS and directly lysed into 1x Laemmli sample buffer containing BME. Each lysate is sonicated and boiled before being tested in Western blot for reactivity to LC3. Tubulin reactivity in each lysate is shown as a loading control.
Neuro2a Chloroquine treated / untreated lysates are provided as positive and negative control for Western blot analysis in Autophagy research. The lysates are provided as one vial of treated and one vial of untreated samples. Use 10 ul per lane for a standard mini-gel blot (approx. 1 mg/ml).
Packaging, Storage & Formulations
Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
1X Laemmli Sample Buffer [31.5 mM Tris HCl, 1% SDS, 10% Glycerol, 0.005% bromophenol blue, pH 6.8]
Lysate Details for Array
Chloroquine Treated / Untreated
- 0.1 ml Neuro2a Chloroquine treated whole cell lysate
- 0.1 ml Neuro2a untreated whole cell lysate
NBP2-49688 contains 0.1ml of Neuro2a Chloroquine treated and 0.1ml of Neuro2a untreated cell lysate
Alternate Names for Neuro2a Chloroquine Treated / Untreated Cell Lysate
- LC3 Lysate
- lc3b autophagy marker
- LC3B western blot
Chloroquine (CQ) or N4-(7-Chloro-4-quinolinyl)-N1,N1-dimethyl-1,4-pentanediamine diphosphate salt (C18H26ClN3. 2H3PO4) is an anti-malarial drug which is well known for its inhibitory effects on apoptosis and autophagy in various cell lines. CQ is a weak base which can partition into the acidic vesicles such as endosomes and lysosomes, thereby, resulting in the inhibition of endosomal acidification and lysosomal enzyme activity. Lysosomotropic autophagy inhibitor CQ decreases the activity of lysosomal acid hydrolase enzymes which impairs the lysosomal function (lysosomal degradation of autophagosomes) ultimately resulting in the accumulation of autophagosomes. Because autophagosomal membranes are rich in lapidated form of LC3 protein (LC3-II), CQ treated cells are often exploited for their usefulness as positive controls in autophagy related research, especially for Western blot and ICC/IF analysis of LC3 of cultured cells. In ICC/IF, the CQ treated cells would be expected to show an enhanced number of autophagosomes compared to vehicle treated cells. Similarly, in WB analysis, the lysate of the CQ treated cells must show an enhanced LC3-II levels compared to the lysate of vehicle treated cells. Neuro2a Chloroquine Treated / Untreated Cell Lysate [NBP2-49688] is provided as a ready to use pair of lysates, 0.1ml of Neuro2a Chloroquine treated (positive control) and 0.1ml of Neuro2a untreated (negative control), for LC3 analysis in Western blot with anti-mouse LC3 antibodies. NBP2-49688 has been validated using monoclonal as well as polyclonal anti-LC3 antibodies, and the lysate of CQ treated cells depicted an increased level of LC3-II (see data image).
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed
for 6 months from date of receipt.
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