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Autophagy Antibody Sampler Kit

Usd: 640

Specification

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Catalog# K2002

Autophagy Antibody Sampler Kit

Application

  • WB

Reactivity

  • Human

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,FC 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

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of LC3B on different lysates with Rabbit anti-LC3B antibody (<a href="/products/ET1701-65" style="font-weight: bold;text-decoration: underline;">ET1701-65</a>) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HeLa treated with 50μM Chloroquine for 18 hours cell lysate<br />Lane 3: C2C12 cell lysate<br />Lane 4: C2C12 treated with 50μM Chloroquine for 18 hours cell lysate<br />Lane 5: C6 cell lysate<br />Lane 6: C6 treated with 50μM Chloroquine for 18 hours cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 14/16 kDa<br />Observed band size: 14/16 kDa<br /><br />Exposure time: 3 minutes;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1701-65" style="font-weight: bold;text-decoration: underline;">ET1701-65</a>) 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 (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ 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 (<a href="/products/ET1611-38" style="font-weight: bold;text-decoration: underline;">ET1611-38</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.<br /><br />Lane 1: NIH/3T3 cell lysate (15 µg/Lane)<br />Lane 2: C2C12 cell lysate (15 µg/Lane)<br />Lane 3: Neuro-2a cell lysate (15 µg/Lane)<br />Lane 4: PC-12 cell lysate (15 µg/Lane)<br />Lane 5: Mouse brain tissue lysate (15 µg/Lane)<br />Lane 6: Rat brain tissue lysate (15 µg/Lane)<br /><br />Predicted band size: 32 kDa<br />Observed band size: 55 kDa<br /><br />Exposure time: 35 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1611-38" style="font-weight: bold;text-decoration: underline;">ET1611-38</a>) 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 (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) 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.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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