LC3 Recombinant antibody

• Featured Product
• KD/KO Validated

LC3 Uni-rAb^TM Recombinant Antibody for WB, IHC, IF/ICC, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat, pig and More (1)

Applications

WB, IHC, IF/ICC, ELISA

Conjugate

Unconjugated

Publications

13

CloneNo.

5P12

Cat no : 81004-1-RR

Synonyms

MAP1LC3B, MAP1A/1BLC3, MAP1 light chain 3-like protein 2, LC3B, Autophagy-related ubiquitin-like modifier LC3 B

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Validation Data Gallery

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  WB analysis using 81004-1-RR

  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.

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  WB analysis of mouse brain using 81004-1-RR

  mouse brain tissue 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.

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  WB analysis of HeLa using 81004-1-RR

  WB result of LC3 antibody (81004-1-RR; 1:20000; room temperature for 1.5 hours) with negative Control and LC3 knockout HeLa cells.

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  IHC staining of mouse brain using 81004-1-RR

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 81004-1-RR (LC3 antibody) at dilution of 1:500 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human colon cancer using 81004-1-RR

  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).

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  IHC staining of human colon cancer using 81004-1-RR

  Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 81004-1-RR (LC3 antibody) at dilution of 1:500 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse brain using 81004-1-RR

  Immunohistochemical analysis of paraffin-embedded mouse brain 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).

• 

  IF Staining of HeLa using 81004-1-RR

  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).

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  Affinity and Kinetic Characterization of 81004-1-RR

  Biolayer interferometry (BLl) kinetic assays of 81004-1-RR against Human LC3 were performed. The affinity constant is 0.967 nM.

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Tested Applications

• Positive WB detected in: mouse brain tissue, HEK-293 cells, HeLa cells, Chloroquine treated HEK-293 cells, Chloroquine treated HeLa cells
• Positive IHC detected in: human colon cancer tissue, mouse brain 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: Chloroquine treated HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:10000
• Immunohistochemistry (IHC): IHC : 1:250-1:1000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:500-1:2000

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 11 publications below
• IHC: See 1 publications below
• IF: See 4 publications below

Product Information

81004-1-RR targets LC3 in WB, IHC, IF/ICC, ELISA applications and shows reactivity with human, mouse, rat, pig samples.

• Tested Reactivity: human, mouse, rat, pig
• Cited Reactivity: human, mouse, rat, bovine
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: LC3 fusion protein Ag6144
• Full Name: microtubule-associated protein 1 light chain 3 beta
• Calculated Molecular Weight: 15 kDa
• Observed Molecular Weight: 14-18 kDa
• GenBank Accession Number: BC067797
• Gene Symbol: LC3B
• Gene ID (NCBI): 81631
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A 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

Map1LC3, also known as LC3, is the human homolog of yeast Atg8 and is involved in the formation of autophagosomal vacuoles, called autophagosomes. Three human Map1LC3 isoforms, MAP1LC3A, MAP1LC3B, and MAP1LC3C, undergo post-translational modifications during autophagy. And they differ in their post-translation modifications during autophagy. Map1LC3 also exists in two modified forms, an 18 kDa cytoplasmic form that was originally identified as a subunit of the microtubule-associated protein 1, and a 14-16 kDa form that is associated with the autophagosome membrane.

Publications

Species	Application	Title
bovine	WB	J Dairy Sci; Subacute ruminal acidosis induces pyroptosis via the mitophagy-mediated NLRP3 inflammasome activation in the livers of dairy cows fed a high-grain diet; Authors - Hongzhu Zhang
human	WB	Discov Oncol; L-cysteine contributes to destructive activities of odontogenic cysts/tumor; Authors - Ji Li
mouse	WB,IF	Int J Mol Sci; Combination of Kaempferol and Docetaxel Induces Autophagy in Prostate Cancer Cells In Vitro and In Vivo; Authors - Qian Zhou
rat		J Hazard Mater; The role of mTORC1/TFEB axis mediated lysosomal biogenesis and autophagy impairment in fluoride neurotoxicity and the intervention effects of resveratrol; Authors - Huayang Tang
mouse	WB	Acta Biomater; Tumor-targeted Delivery of Copper-manganese Biomineralized Oncolytic Adenovirus for Colorectal Cancer Immunotherapy; Authors - Yi-Shu Li
human	WB	Front Pharmacol; Inhibition of NNMT enhances drug sensitivity in lung cancer cells through mediation of autophagy; Authors - Jian Wang