LC3 Polyclonal antibody

LC3 Polyclonal Antibody for WB, IF, IHC, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat, zebrafish and More (3)

Applications

WB, IHC, IF/ICC, ELISA

Conjugate

Unconjugated

Publications

70

Cat no : 12135-1-AP

Synonyms

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Validation Data Gallery

• 

  WB analysis of HeLa using 12135-1-AP

  Chloroquine treated HeLa cells were subjected to SDS PAGE followed by western blot with 12135-1-AP (LC3 Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HEK-293 using 12135-1-AP

  Chloroquine treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 12135-1-AP (LC3 Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  IHC staining of human breast cancer using 12135-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 12135-1-AP (LC3 antibody) at dilution of 1:200 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human breast cancer using 12135-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 12135-1-AP (LC3 antibody) at dilution of 1:200 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human liver cancer using 12135-1-AP

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 12135-1-AP (LC3 antibody) at dilution of 1:200 (under 10x lens).

• 

  IHC staining of human liver cancer using 12135-1-AP

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 12135-1-AP (LC3 antibody) at dilution of 1:200 (under 40x lens).

• 

  IF Staining of HEK-293 using 12135-1-AP

  Immunofluorescent analysis of starvation treated HEK-293 cells using 12135-1-AP (LC3 Antibody) at dilution of 1:50 and Alexa Fluor 488-conjugated Goat Anti-Rabbit IgG(H+L). Cells were fixed with ethanol at -20 °C. 20 mM chloroquine was used to block the autophagy flux.

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Tested Applications

• Positive WB detected in: Chloroquine treated HEK-293 cells, Chloroquine treated HeLa cells
• Positive IHC detected in: human breast cancer tissue, human liver cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: Starvation treated HEK-293 cells

Recommended dilution

• Western Blot (WB): WB : 1:500-1:2000
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 58 publications below
• IHC: See 7 publications below
• IF: See 22 publications below

Product Information

12135-1-AP targets LC3 in WB, IHC, IF/ICC, ELISA applications and shows reactivity with human, mouse, rat, zebrafish samples.

• Tested Reactivity: human, mouse, rat, zebrafish
• Cited Reactivity: human, mouse, rat, pig, chicken, turtle
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: LC3 fusion protein Ag2781
• Full Name: microtubule-associated protein 1 light chain 3 alpha
• Calculated Molecular Weight: 121 aa, 14 kDa
• GenBank Accession Number: BC015810
• Gene Symbol: LC3
• Gene ID (NCBI): 84557
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

LC3, also known as microtubule-associated protein 1 light chain 3 (MAP1LC3), is a mammalian homolog of the yeast Atg8p (autophagy-related 8 protein) that is involved in autophagy process. Three isoforms of LC3 exist as LC3A,LC3B, and LC3C. LC3 is first synthesized as pro-LC3, and undergoes C-terminal cleavage to yield a cytosolic precursor known as LC3-I. During autophagy LC3-I is subsequently conjugated to PE lipid as lipidated form: LC3-II. The lipidated form migrates faster on SDS-PAGE gel. The increased the level of LC3-II has been widely used as autophagy marker. This antibody recognizes both LC3-I (18-20 kDa) and LC3-II (14-16 kDa).

Publications

Species	Application	Title
mouse	IF,WB	Autophagy; Analysis of ATG4C function in vivo; Authors - Isaac Tamargo-Gómez
mouse	WB	J Clin Invest; Autophagy is essential for mouse sense of balance.; Authors - Mari?o Guillermo G
mouse	WB	Autophagy; Assessing autophagy in the context of photodynamic therapy.; Authors - Reiners John J JJ Jr
mouse	WB	Cell Death Differ; ATG4D is the main ATG8 delipidating enzyme in mammalian cells and protects against cerebellar neurodegeneration.; Authors - Isaac Tamargo-Gómez
human	WB	EMBO J; Unsaturated fatty acids induce non-canonical autophagy.; Authors - Mireia Niso-Santano
human	WB	Oxid Med Cell Longev; Enhanced p62-NRF2 Feedback Loop due to Impaired Autophagic Flux Contributes to Arsenic-Induced Malignant Transformation of Human Keratinocytes.; Authors - Xiafang Wu

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