HeLa Chloroquine Treated / Untreated Cell Lysate

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Western Blot: HeLa Chloroquine Treated / Untreated Cell Lysate [NBP2-49689] - HeLa cells were treated with (+) or without 50 uM (-) of Chloriquine (CQ) for 24 hours. Total cell lysates were prepared and separated on a ...read more
Western Blot: HeLa Chloroquine Treated / Untreated Cell Lysate [NBP2-49689] - 10 ug of Chloroquine treated (+) and untreated (-) HeLa lysates in 1x Laemmli sample buffer (NBP2-49689) were separated on a 4-15% gel by ...read more
Western Blot: HeLa Chloroquine Treated / Untreated Cell Lysate [NBP2-49689] - Human Cervical carcinoma cells (HeLa) were treated with (+) and without (-) 50 uM Chloroquine overnight. Whole cell protein lysates were ...read more
Western Blot: HeLa Chloroquine Treated / Untreated Cell Lysate [NBP2-49689] - HeLa cells were treated with vehicle (-) or with 50 uM chloriquine (+) for 24 hours and the total cell lysates were prepared. The lysates ...read more

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB, PAGE
Concentration
1.0 mg/ml

Order Details

HeLa Chloroquine Treated / Untreated Cell Lysate Summary

Description
NBP2-49689 contains 1 vial each of 0.1ml of HeLa Chloroquine treated and 0.1ml of HeLa untreated cell lysate. This lysate set is useful as a positive and negative controls for targets such as LC3, p62 etc. and the researchers should use anti-Human primary antibodies when working with these lysates.
Marker
Autophagy/LC3 Positive Control
Preparation
Method
HeLa cells were cultured under standard laboratory conditions 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.

Applications/Dilutions

Application Notes
HeLa Chloroquine treated / untreated whole cell 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

Storage
Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
Buffer
The protein lysate is prepared in 1x Laemmli sample buffer with BME. Boiling before loading is not necessary.
Preservative
No Preservative
Concentration
1.0 mg/ml

Lysate Details for Array

Type
Cell
Tissue
Cervix
Subcellular Fraction
Chloroquine Treated / Untreated

Kit Components

Components
  1. 0.1 ml HeLa Chloroquine treated whole cell lysate
  2. 0.1 ml HeLa untreated whole cell lysate

Notes

NBP2-49689 contains 0.1ml of HeLa Chloroquine treated and 0.1ml of HeLa untreated cell lysate

Alternate Names for HeLa Chloroquine Treated / Untreated Cell Lysate

  • LC3 Lysate
  • LC3B western blot

Background

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. HeLa Chloroquine Treated / Untreated Cell Lysate [NBP2-49689] is provided as a ready to use pair of lysates, 0.1ml of HeLa Chloroquine treated (positive control) and 0.1ml of HeLa untreated (negative control), for Western blot with anti-Human LC3 or p62 antibodies. NBP2-49689 has been validated using monoclonal as well as polyclonal anti-LC3 and antibodies, and the lysate of CQ treated cells depicted an increased level of LC3-II (see data image).

Limitations

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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The Role of LC3 within the Autophagic Pathway
We at Novus Biologicals have a broad antibody database covering the area of autophagy - over 1400 reagents in total. Autophagy is the bulk degradation of cytoplasmic components - literally, self-digestion of the cell. Double-membrane vesicles, called...  Read full blog post.

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