Autophagy Antibody Sampler Kit
Catalog# K2002
Autophagy Antibody Sampler Kit
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WB
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Human
General Overview
Kit Components
| Product Includes | Specification | Application | Reactivity | Mw |
|---|---|---|---|---|
| LC3B[ET1701-65] | 20µl | WB,IF-Cell,IHC-P,IF-Tissue,IP,mIHC | Human,Mouse,Rat | Predicted band size: 14/16 kDa |
| ATG5[ET1611-38] | 20µl | WB,IF-Cell,IF-Tissue,IHC-P,IP,FC | Human,Mouse,Rat,Monkey | Predicted band size: 32 kDa |
| ATG12[HA721504] | 20µl | WB,IF-Cell,FC | Human | Predicted band size: 15 kDa |
| ATG16L1[ET7106-65] | 20µl | WB,IHC-P | Human,Mouse,Rat | Predicted band size: 68 kDa |
| ATG7[ET1610-53] | 20µl | WB,IF-Tissue,IHC-P,IP,FC | Human,Rat | Predicted band size: 78 kDa |
| Apg3[ET1706-29] | 20µl | WB,IHC-P | Human,Mouse,Rat | 40 kDa |
| Beclin 1[HA721216] | 20µl | WB,IHC-P,IF-Cell | Human,Mouse,Rat | Predicted band size: 52 kDa |
| Alpaca anti-Rabbit IgG Fc, Recombinant VHH[HA1031] | 100µl | IP,ELISA,IHC-P,WB | Rabbit |
Product Description
The Autophagy Antibody Sampler Kit provides an economical means to investigate the molecular machinery of autophagy within the cell. The kit contains enough primary and secondary antibodies to perform two Western blot experiments.
Product Features
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Background
Autophagy is a catabolic process for the autophagosomic-lysosomal degradation of bulk cytoplasmic contents. Autophagy is generally activated by conditions of nutrient deprivation but has also been associated with a number of physiological processes including development, differentiation, neurodegeneration, infection, and cancer. </br>The molecular machinery of autophagy was largely discovered in yeast and referred to as autophagy-related (Atg) genes. Formation of the autophagosome involves a ubiquitin-like conjugation system in which Atg12 is covalently bound to Atg5 and targeted to autophagosome vesicles. This conjugation reaction is mediated by the ubiquitin E1-like enzyme Atg7 and the E2-like enzyme Atg10.
Data Links
Background References
1. Reggiori, F. and Klionsky, D.J. (2002) Eukaryot Cell 1, 11-21.
2. Codogno, P. and Meijer, A.J. (2005) Cell Death Differ 12 Suppl 2, 1509-18.
3. Levine, B. and Yuan, J. (2005) J Clin Invest 115, 2679-88.
4. Mizushima, N. et al. (1998) J Biol Chem 273, 33889-92.
5. Mizushima, N. et al. (1998) Nature 395, 395-8.
6. Suzuki, K. et al. (2001) EMBO J 20, 5971-81.
7. Tanida, I. et al. (1999) Mol Biol Cell 10, 1367-79.
8. Shintani, T. et al. (1999) EMBO J 18, 5234-41.
Images
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☑ Cell treatment (CT)
Western blot analysis of LC3B on different lysates with Rabbit anti-LC3B antibody (ET1701-65) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: HeLa treated with 50μM Chloroquine for 18 hours cell lysate
Lane 3: C2C12 cell lysate
Lane 4: C2C12 treated with 50μM Chloroquine for 18 hours cell lysate
Lane 5: C6 cell lysate
Lane 6: C6 treated with 50μM Chloroquine for 18 hours cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 14/16 kDa
Observed band size: 14/16 kDa
Exposure time: 3 minutes;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1701-65) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Western blot analysis of ATG5 on different lysates with Rabbit anti-ATG5 antibody (ET1611-38) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.
Lane 1: NIH/3T3 cell lysate (15 µg/Lane)
Lane 2: C2C12 cell lysate (15 µg/Lane)
Lane 3: Neuro-2a cell lysate (15 µg/Lane)
Lane 4: PC-12 cell lysate (15 µg/Lane)
Lane 5: Mouse brain tissue lysate (15 µg/Lane)
Lane 6: Rat brain tissue lysate (15 µg/Lane)
Predicted band size: 32 kDa
Observed band size: 55 kDa
Exposure time: 35 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1611-38) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
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