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Autophagy Expanded Antibody Kit

Host

Rabbit/Mouse

Reactivity

Target dependent

Applications

Target dependent

Conjugate

Unconjugated

Cat no : PK30005

Validation Data Gallery

Applications:
  • Various lysates were subjected to SDS PAGE followed by western blot with 20986-1-AP (ULK1 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

    Various lysates were subjected to SDS PAGE followed by western blot with 20986-1-AP (ULK1 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

  • Various lysates were subjected to SDS PAGE followed by western blot with 66665-1-Ig (Beclin 1 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

    Various lysates were subjected to SDS PAGE followed by western blot with 66665-1-Ig (Beclin 1 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

  • mouse brain tissue were subjected to SDS PAGE followed by western blot with 26276-1-AP (ATG9A Antibody) at dilution of 1:600 incubated at room temperature for 1.5 hours.

    mouse brain tissue were subjected to SDS PAGE followed by western blot with 26276-1-AP (ATG9A Antibody) at dilution of 1:600 incubated at room temperature for 1.5 hours.

  • Untreated and chloroquine treated HEK-293 cells, untreated and chloroquine treated HeLa cells were subjected to SDS PAGE followed by western blot with 81004-1-RR (LC3 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

    Untreated and chloroquine treated HEK-293 cells, untreated and chloroquine treated HeLa cells were subjected to SDS PAGE followed by western blot with 81004-1-RR (LC3 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

  • Lysates of HepG2 cells treated with Chloroquine or not were subjected to SDS PAGE followed by western blot with 80294-1-RR (P62,SQSTM1 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

    Lysates of HepG2 cells treated with Chloroquine or not were subjected to SDS PAGE followed by western blot with 80294-1-RR (P62,SQSTM1 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

  • Various lysates were subjected to SDS PAGE followed by western blot with 81803-1-RR (ATG5 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

    Various lysates were subjected to SDS PAGE followed by western blot with 81803-1-RR (ATG5 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

  • Various lysates were subjected to SDS PAGE followed by western blot with 29445-1-AP (ATG16L1 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

    Various lysates were subjected to SDS PAGE followed by western blot with 29445-1-AP (ATG16L1 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

  • Various lysates were subjected to SDS PAGE followed by western blot with 11264-1-AP (ATG12 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

    Various lysates were subjected to SDS PAGE followed by western blot with 11264-1-AP (ATG12 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

  • Various lysates were subjected to SDS PAGE followed by western blot with 80992-1-RR (ubiquitin antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

    Various lysates were subjected to SDS PAGE followed by western blot with 80992-1-RR (ubiquitin antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

  • U-937 cells were subjected to SDS PAGE followed by western blot with 21997-1-AP (LAMP1 antibody at dilution of 1:2000 incubated at room temperature for 1.5 hours.

    U-937 cells were subjected to SDS PAGE followed by western blot with 21997-1-AP (LAMP1 antibody at dilution of 1:2000 incubated at room temperature for 1.5 hours.

  • IP result of anti-P62,SQSTM1(IP:80294-1-RR, 4ug; Detection:80294-1-RR 1:500) with HEK-293 cells lysate 1640 ug.

    IP result of anti-P62,SQSTM1(IP:80294-1-RR, 4ug; Detection:80294-1-RR 1:500) with HEK-293 cells lysate 1640 ug.

  • IP result of anti-ATG5(IP:81803-1-RR, 4ug; Detection:81803-1-RR 1:2000) with HeLa cells lysate 1800 ug.

    IP result of anti-ATG5(IP:81803-1-RR, 4ug; Detection:81803-1-RR 1:2000) with HeLa cells lysate 1800 ug.

  • Immunohistochemical analysis of paraffin-embedded human stomach tissue slide using 66665-1-Ig (Beclin 1 antibody) at dilution of 1:500 (under 10x lens).

    Immunohistochemical analysis of paraffin-embedded human stomach tissue slide using 66665-1-Ig (Beclin 1 antibody) at dilution of 1:500 (under 10x lens).

  • Immunohistochemical analysis of paraffin-embedded human heart tissue slide using 26276-1-AP (ATG9A Antibody) at dilution of 1:100 (under 10x lens).

    Immunohistochemical analysis of paraffin-embedded human heart tissue slide using 26276-1-AP (ATG9A Antibody) at dilution of 1:100 (under 10x lens).

  • Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 81004-1-RR (LC3 antibody) at dilution of 1:500 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

    Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 81004-1-RR (LC3 antibody) at dilution of 1:500 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded human pancreas cancer tissue slide using 80294-1-RR (P62,SQSTM1 antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

    Immunohistochemical analysis of paraffin-embedded human pancreas cancer tissue slide using 80294-1-RR (P62,SQSTM1 antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded human ovary tumor tissue slide using 81803-1-RR (ATG5 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

    Immunohistochemical analysis of paraffin-embedded human ovary tumor tissue slide using 81803-1-RR (ATG5 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 29445-1-AP (ATG16L1 antibody) at dilution of 1:400 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

    Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 29445-1-AP (ATG16L1 antibody) at dilution of 1:400 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 11264-1-AP (ATG12 antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

    Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 11264-1-AP (ATG12 antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded human pancreas cancer tissue slide using 80992-1-RR (ubiquitin antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

    Immunohistochemical analysis of paraffin-embedded human pancreas cancer tissue slide using 80992-1-RR (ubiquitin antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 21997-1-AP (LAMP1 antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

    Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 21997-1-AP (LAMP1 antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

  • Immunofluorescent analysis of (4% PFA) fixed mouse heart tissue using 66665-1-Ig (Beclin 1 antibody) at dilution of 1:100 and Alexa Fluor 488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

    Immunofluorescent analysis of (4% PFA) fixed mouse heart tissue using 66665-1-Ig (Beclin 1 antibody) at dilution of 1:100 and Alexa Fluor 488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

  • Immunofluorescent analysis of (4% PFA) fixed mouse brain tissue using 26276-1-AP (ATG9A antibody) at dilution of 1:50 and Alexa Fluor 488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

    Immunofluorescent analysis of (4% PFA) fixed mouse brain tissue using 26276-1-AP (ATG9A antibody) at dilution of 1:50 and Alexa Fluor 488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

  • Immunofluorescent analysis of (-20°C Methanol) fixed Chloroquine treated HeLa cells using LC3 antibody (81004-1-RR, Clone: 5P12 ) at dilution of 1:1000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CoraLite®594 Beta Actin antibody (CL594-66009, Clone: 2D4H5, red).

    Immunofluorescent analysis of (-20°C Methanol) fixed Chloroquine treated HeLa cells using LC3 antibody (81004-1-RR, Clone: 5P12 ) at dilution of 1:1000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CoraLite®594 Beta Actin antibody (CL594-66009, Clone: 2D4H5, red).

  • Immunofluorescent analysis of (-20°C Ethanol) fixed Chloroquine treated HeLa cells using P62,SQSTM1 antibody (80294-1-RR, Clone: 1D17 ) at dilution of 1:500 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CL594-Phalloidin (red).

    Immunofluorescent analysis of (-20°C Ethanol) fixed Chloroquine treated HeLa cells using P62,SQSTM1 antibody (80294-1-RR, Clone: 1D17 ) at dilution of 1:500 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CL594-Phalloidin (red).

  • Immunofluorescent analysis of (-20°C Ethanol) fixed HEK-293 cells using ATG16L1 antibody (29445-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CL594-Phalloidin (red).

    Immunofluorescent analysis of (-20°C Ethanol) fixed HEK-293 cells using ATG16L1 antibody (29445-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CL594-Phalloidin (red).

  • Immunofluorescent analysis of (4% PFA) fixed HeLa cells using ubiquitin antibody (80992-1-RR, Clone: 6H6 ) at dilution of 1:1000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CL594-phalloidin (red).

    Immunofluorescent analysis of (4% PFA) fixed HeLa cells using ubiquitin antibody (80992-1-RR, Clone: 6H6 ) at dilution of 1:1000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CL594-phalloidin (red).

Product Information

The Autophagy Expanded Antibody Kit provides a cost-effective tool for studying key proteins involved in the autophagy pathway. Perfect for researchers starting a new project, screening multiple prospective targets or those who simply require less volume.

Kit Components

The Autophagy Expanded Antibody Kit contains antibodies against 10 key protein targets playing critical roles in the autophagy pathway. 

AntigenCatalog No.Host, clonalityTested ReactivityApplicationsVolume
ULK120986-1-AP            Rabbit polyclonalH, M, RWB, IHC, IF, ELISA20 uL
Beclin 166665-1-Ig            Mouse monoclonalH, M, RWB, IHC, IF, ELISA20 uL
ATG9A26276-1-AP            Rabbit polyclonalH, M, RWB, IHC, IF, ELISA20 uL
LC381004-1-RR            Rabbit monoclonalH, M, R, PgWB, IHC, IF, ELISA20 uL
p6280294-1-RR            Rabbit monoclonalHWB, IP, IHC, IF, ELISA20 uL
ATG581803-1-RR            Rabbit monoclonalH, M, RWB, IP, IHC, ELISA20 uL
ATG16 L129445-1-AP            Rabbit polyclonalH, M, RWB, IHC, IF, ELISA20 uL
ATG1211264-1-AP            Rabbit polyclonalH, MWB, IHC, IF, ELISA20 uL
Ubiquitin80992-1-RR            Rabbit monoclonalH, M, RWB, IHC, IF, ELISA20 uL
LAMP121997-1-AP            Rabbit polyclonalHWB, IP, IHC, FC, ELISA20 uL

Also see our 'Autophagy Essentials Antibody Kit' on the following page  https://www.ptglab.com/products/Autophagy-Essentials-Antibody-Kit-PK30004.htm

Storage

Store at -20°C. Stable for one year from the date of receipt.

Background Information

Autophagy is a highly dynamic process consisting of the following three steps: (1) autophagosome formation, (2) autophagosome-lysosome fusion, and (3) degradation. It can be induced by multiple signaling pathways related to various triggers including nutrient deprivation, growth factor signaling, and cellular stress. ULK1 and Beclin 1 are critical for the initiation of autophagy. The process of autophagosome formation proceeds through the steps of initiation, nucleation, elongation, closure, and ultimately fusion, each of which is regulated by various ATG proteins. Ubiquitination of various autophagy-related proteins and regulatory proteins are critical for the precise regulation of the autophagy pathway.  

The ideal approach for measuring autophagy is to assess autophagic flux, which represents the rate of degradation of the autophagic pathway. The most widely used method for measuring autophagic flux is to detect the processing of the autophagosomal membrane protein, LC3. Analyzing autophagy substrates such as p62/SQSTM1 is often recommended in addition to measuring LC3-II turnover for accurate assessment of autophagic flux. The fusion of autophagosomes with lysosomes can be monitored by analyzing the autophagosomal marker LC3 and the lysosomal marker, LAMP simultaneously. 

Standard Protocols

Click here to view our standard protocols for various applications including WB, IP, IHC, IF, FC, and ELISA.

Cited in Article as

PK30005, Autophagy Expanded Antibody Kit, Proteintech, IL, USA