LC3 Polyclonal antibody

• Featured Product
• KD/KO Validated

LC3 Polyclonal Antibody for FC, IF, IHC, IP, WB, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat and More (7)

Applications

WB, IP, IHC, IF, FC, ELISA

Conjugate

Unconjugated

Publications

548

Reviews (7)

Cat no : 14600-1-AP

Synonyms

LC3, LC3B, MAP1A/1BLC3, MAP1A/MAP1B LC3 B, MAP1A/MAP1B light chain 3 B, MAP1ALC3, MAP1LC3B

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Validation Data Gallery

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  WB analysis using 14600-1-AP

  Chloroquine treated HepG2 cells (left) and untreated HepG2 cells (right) after serum deprived for 2 hours were subjected to SDS PAGE followed by western blot with 14600-1-AP (LC3 antibody) at dilution of 1:2500 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HepG2 using 14600-1-AP

  WB result of LC3 antibody (14600-1-AP; 1:6000; incubated at room temperature for 1.5 hours) with sh-Control and sh-LC3 transfected HepG2 cells.

• 

  WB analysis of HEK-293 using 14600-1-AP

  CQ treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 14600-1-AP (LC3 Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HeLa using 14600-1-AP

  HeLa cells were subjected to SDS PAGE followed by western blot with 14600-1-AP (LC3 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of rat brain using 14600-1-AP

  rat brain tissue were subjected to SDS PAGE followed by western blot with 14600-1-AP (LC3 Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of mouse testis using 14600-1-AP

  mouse testis tissue were subjected to SDS PAGE followed by western blot with 14600-1-AP (LC3 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

• 

  WB analysis of human skeletal muscle using 14600-1-AP

  human skeletal muscle tissue were subjected to SDS PAGE followed by western blot with 14600-1-AP (LC3 antibody) at dilution of 1:300 incubated at room temperature for 1.5 hours.

• 

  WB analysis of mouse brain using 14600-1-AP

  mouse brain tissue were subjected to SDS PAGE followed by western blot with 14600-1-AP (LC3 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HSC-T6 using 14600-1-AP

  HSC-T6 cells were subjected to SDS PAGE followed by western blot with 14600-1-AP (LC3 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of mouse lung using 14600-1-AP

  mouse lung tissue were subjected to SDS PAGE followed by western blot with 14600-1-AP (LC3 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

• 

  WB analysis of NIH/3T3 using 14600-1-AP

  NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 14600-1-AP (LC3 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

• 

  WB analysis of mouse heart using 14600-1-AP

  mouse heart tissue were subjected to SDS PAGE followed by western blot with 14600-1-AP (LC3 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

• 

  WB analysis of mouse liver using 14600-1-AP

  mouse liver tissue were subjected to SDS PAGE followed by western blot with 14600-1-AP (LC3 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

• 

  IP experiment of mouse brain using 14600-1-AP

  IP Result of anti-LC3 (IP:14600-1-AP, 3ug; Detection:14600-1-AP 1:1000) with mouse brain tissue lysate 4000ug.

• 

  IHC staining of human gliomas using 14600-1-AP

  Immunohistochemical analysis of paraffin-embedded human gliomas tissue slide using 14600-1-AP (LC3 antibody) at dilution of 1:600 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human gliomas using 14600-1-AP

  Immunohistochemical analysis of paraffin-embedded human gliomas tissue slide using 14600-1-AP (LC3 antibody) at dilution of 1:600 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human testis using 14600-1-AP

  Immunohistochemical analysis of paraffin-embedded human testis using 14600-1-AP (LC3 antibody) at dilution of 1:50 (under 10x lens).

• 

  IHC staining of human testis using 14600-1-AP

  Immunohistochemical analysis of paraffin-embedded human testis using 14600-1-AP (LC3 antibody) at dilution of 1:50 (under 40x lens).

• 

  IHC staining of human breast cancer using 14600-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 14600-1-AP (LC3 Antibody) at dilution of 1:50 (under 10x lens).

• 

  IHC staining of human breast cancer using 14600-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 14600-1-AP (LC3 Antibody) at dilution of 1:50 (under 40x lens).

• 

  IHC staining of mouse brain using 14600-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 14600-1-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 mouse brain using 14600-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 14600-1-AP (LC3 antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of HeLa using 14600-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed Chloroquine treated HeLa cells using LC3 antibody (14600-1-AP) at dilution of 1:500 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

• 

  IF Staining of U2OS using 14600-1-AP

  Immunofluorescent analysis of (-20°C Methanol) fixed Chloroquine treated U2OS cells using LC3 antibody (14600-1-AP) at dilution of 1:500 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

• 

  IF Staining of HepG2 using 14600-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed CQ treated HepG2 cells using 14600-1-AP (LC3 antibody) at dilution of 1:100 and CoraLite488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

• 

  IF Staining of HepG2 using 14600-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed CQ treated HepG2 cells using 14600-1-AP (LC3 antibody) at dilution of 1:200 and CoraLite488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

• 

  FC experiment of HeLa using 14600-1-AP

  1X10^6 HeLa cells were stained with 0.2ug LC3 antibody (14600-1-AP, red) and control antibody (blue). Fixed with 90% MeOH blocked with 3% BSA (30 min). Alexa Fluor 488-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) with dilution 1:1000.

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Tested Applications

• Positive WB detected in: Chloroquine treated HepG2 cells, HepG2 cells, mouse brain tissue, mouse lung tissue, HSC-T6 cells, mouse testis tissue, human skeletal muscle tissue, NIH/3T3 cells, Chloroquine treated HEK-293 cells, rat brain tissue, mouse heart tissue, mouse liver tissue, HeLa cells
• Positive IP detected in: mouse brain tissue
• Positive IHC detected in: human gliomas tissue, mouse brain tissue, human testis tissue, human breast 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 detected in: Chloroquine treated HeLa cells, Chloroquine treated HepG2 cells, Chloroquine treated U2OS cells
• Positive FC detected in: HeLa cells

Planning an IF experiment? We recommend our CoraLite®488, CoraLite®594 and CoraLite®647 conjugated versions of this antibody.

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:5000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for IP and 1:500-1:2000 for WB
• Immunohistochemistry (IHC): IHC : 1:300-1:1200
• Immunofluorescence (IF): IF : 1:250-1:1000

Sample-dependent, check data in validation data gallery

Published Applications

• WB: See 483 publications below
• IHC: See 56 publications below
• IF: See 134 publications below
• IP: See 1 publications below

Product Information

14600-1-AP targets LC3 in WB, IP, IHC, IF, FC, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat, bovine, chicken, duck, goat, monkey, pig, rabbit
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• 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: MAP1LC3B
• Gene ID (NCBI): 81631
• 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.

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. This antibody can cross react with MAP1LC3A, MAP1LC3B, and MAP1LC3C.

Publications

Species	Application	Title
human	WB	Nanotoxicology; Silver nanoparticles induce protective autophagy via Ca2+/CaMKKβ/AMPK/mTOR pathway in SH-SY5Y cells and rat brains.; Authors - Lin Li
human	WB	J Cancer; Lyn Kinase Promotes the Proliferation of Malignant Melanoma Cells through Inhibition of Apoptosis and Autophagy via the PI3K/Akt Signaling Pathway.; Authors - Qianqian Zhang
mouse	WB	Int Immunopharmacol; Protective effects of levo-tetrahydropalmatine on hepatic ischemia/reperfusion injury are mediated by inhibition of the ERK/NF-κB pathway.; Authors - Qiang Yu
mouse	IF	Connect Tissue Res; Rapamycin attenuates articular cartilage degeneration by inhibiting β-catenin in a murine model of osteoarthritis.; Authors - Long Ma
human	WB	Cell Death Dis; Microglia as modulators of exosomal alpha-synuclein transmission.; Authors - Yun Xia
rat	WB	FASEB J; STAT3 phosphorylation mediates high glucose-impaired cell autophagy in an HDAC1-dependent and -independent manner in Schwann cells of diabetic peripheral neuropathy.; Authors - Wei Du

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive forproviding their feedback.

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FH

Zhihao (Verified Customer) (07-25-2022)

We could detect one form of LC3 in our cell samples, which should be two.

• Applications: Western Blot
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: GFP-Parkin transfected HeLa cell

LC3 Antibody Western Blot validation (1:1000 dilution) in GFP-Parkin transfected HeLa cell (Cat no:14600-1-AP)

FH

S (Verified Customer) (12-31-2021)

I have tried three different LC3B antibodies from three different companies so far. This antibody is by far the best (cost efficient as well)

• Applications: Western Blot
• Primary Antibody Dilution: 1:2000
• Cell Tissue Type: HCT116

LC3 Antibody Western Blot validation (1:2000 dilution) in HCT116 (Cat no:14600-1-AP)

FH

YING (Verified Customer) (09-25-2021)

The antibody works very well. Distinguished LC3-I and LC3-II bands. An increase of LC3-II level in response to treatment with bafilomycin A.

• Applications: Western Blot
• Primary Antibody Dilution: 1:1500
• Cell Tissue Type: mouse cells

LC3 Antibody Western Blot validation (1:1500 dilution) in mouse cells (Cat no:14600-1-AP)

FH

Eric (Verified Customer) (03-13-2021)

Worked very well. I was able to get the LC3A/B bands on western blot using a 12.5% gel and prominent bands on a 8% gel. IF imaging worked fairly well, I used 1:100 which was likely too low so I will repeat at higher but got signal nonetheless. This performed better than some of the other LC3 antibodies we tested.

• Applications: Western Blot, Immunofluorescence
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: Microglia

LC3 Antibody Western Blot,Immunofluorescence validation (1:1000 dilution) in Microglia (Cat no:14600-1-AP)

FH

Elzbieta (Verified Customer) (09-23-2020)

The antibody works very nicely in WB. Clear signal, no aspecific bands. Our lab discarded other products and switched to this antibody.

• Applications: Western Blot
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: mouse liver, human HepG2 cells

FH

An (Verified Customer) (09-17-2020)

Used for IHC on zebrafish cryosections of eyes. Worked well.

• Applications: Immunohistochemistry

FH

David (Verified Customer) (03-30-2020)

Very nice visualisation of LC3-I and LC3-II. Run on 20% gel and significant changes in response to treatment with bafilomycin A and concanamycin A.

• Applications: Western Blot
• Primary Antibody Dilution: 1:500
• Cell Tissue Type: SH-SY5Y

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