![Western Blot: Neuro2a Chloroquine Treated / Untreated Cell Lysate [NBP2-49688] - Mouse Neuoblast cells (Neuro2A) were treated with (+) and without (-) 50 uM Chloroquine overnight. Whole cell protein lysates were prepared in 1x Laemmli sample buffer and approximately 10 ug of each lysate (NBP2-49688) was separated on a 4-15% gel by SDS-PAGE, transferred to 0.2 um PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 1 ug/ml anti-LC3 (NB100-2220) and 1 ug/ml anti-alpha tubulin (NB100-690) as a loading control, and detected with the appropriate secondary antibodies using chemiluminescence.](http://images.novusbio.com/fullsize/Neuro2a-Chloroquine-Treated---Untreated-Cell-Lysate-Western-Blot-NBP2-49688-img0005.jpg "Western Blot: Neuro2a Chloroquine Treated / Untreated Cell Lysate [NBP2-49688] - Mouse Neuoblast cells (Neuro2A) were treated with (+) and without (-) 50 uM Chloroquine overnight. Whole cell protein lysates were prepared in 1x Laemmli sample buffer and approximately 10 ug of each lysate (NBP2-49688) was separated on a 4-15% gel by SDS-PAGE, transferred to 0.2 um PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 1 ug/ml anti-LC3 (NB100-2220) and 1 ug/ml anti-alpha tubulin (NB100-690) as a loading control, and detected with the appropriate secondary antibodies using chemiluminescence.")
| Reactivity | MuSpecies Glossary |
| Applications | WB, PAGE |
| Concentration | 1.0 mg/ml |
| Description | 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. |
| Marker | Autophagy/LC3 Positive Control |
| Preparation Method |
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. |
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| Application Notes | 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). |
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| Storage | Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles. |
| Buffer | 1X Laemmli Sample Buffer [31.5 mM Tris HCl, 1% SDS, 10% Glycerol, 0.005% bromophenol blue, pH 6.8] |
| Preservative | No Preservative |
| Concentration | 1.0 mg/ml |
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| Publication using NBP2-49688 | Applications | Species |
|---|---|---|
| Cachon-Gonzalez Mb, Wang S, Cox Tm Expression of Ripk1 and DAM genes correlates with severity and progression of Krabbe disease Human molecular genetics 2021-06-25 [PMID: 34172992] (WB) | WB |
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![Western Blot: Neuro2a Chloroquine Treated / Untreated Cell Lysate [NBP2-49688] - Western Blot: [NBP2-49688] - 10 ug of Chloroquine treated (+) and untreated (-) Neuro2A lysates in 1x Laemmli sample buffer (NBP2-49688) were separated on a 4-15% gel by SDS-PAGE, transferred to 0.2 um PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 1 ug/ml monoclonal anti-LC3 (NBP2--46892) and detected with an anti-rabbit secondary antibody using chemiluminescence.](http://images.novusbio.com/fullsize/Neuro2a-Chloroquine-Treated---Untreated-Cell-Lysate-Western-Blot-NBP2-49688-img0004.jpg "Western Blot: Neuro2a Chloroquine Treated / Untreated Cell Lysate [NBP2-49688] - Western Blot: [NBP2-49688] - 10 ug of Chloroquine treated (+) and untreated (-) Neuro2A lysates in 1x Laemmli sample buffer (NBP2-49688) were separated on a 4-15% gel by SDS-PAGE, transferred to 0.2 um PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 1 ug/ml monoclonal anti-LC3 (NBP2--46892) and detected with an anti-rabbit secondary antibody using chemiluminescence.")