PRODUCT
Summary
LC3B antibody recognizes LC3B, an autophagosomal membrane structural protein with a predicted molecular weight of 14.7 kDa. LC3B protein is a member of the highly conserved Atg8 protein family, which encompasses the LC3 and GABARAP subfamilies. The key function of LC3B involves autophagy substrate selection during autophagosome formation. Therefore, LC3B antibody is of great utility for the study of autophagosome biogenesis.
APPLICATION
Application Note
*Optimal dilutions/concentrations should be determined by the researcher.
| Application |
Recommended Dilution |
| 1:500-1:3000 |
| 1:100-1:1000 |
| 1:100-1:1000 |
| Assay dependent |
| 1:50-1:200 |
| 1:100-1:500 |
| Assay dependent |
Not tested in other applications.
Calculated MW
Positive Control
Huh7 (HCV-infected) , Huh7 (3uM-Thapsigargin treatment for 12hr) , mouse brain , Rat brain , NT2D1 , PC-3 , U87-MG , SK-N-SH , HepG2 (3 μM Thapsigargin treatment for 16hr) , HepG2 (3 μM Thapsigargin treatment for 24hr) , HeLa (50μM Chloroquine treatment for 24 hr)
Product Note
Cross-reactivity may occur with other LC3 isoforms
Predict Reactivity
Zebrafish, Bovine, Chicken, Japanese Medaka, Xenopus tropicalis, Chimpanzee, Rhesus Monkey(>80% identity)
PROPERTIES
Form
Liquid
Buffer
PBS, 20% Glycerol
Preservative
0.025% ProClin 300
Storage
Store as concentrated solution. Centrifuge briefly prior to opening vial. For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, aliquot and store at -20ºC or below. Avoid multiple freeze-thaw cycles.
Concentration
0.34 mg/ml (Please refer to the vial label for the specific concentration.)
Antigen Species
Human
Immunogen
Carrier-protein conjugated synthetic peptide encompassing a sequence within the N-terminus region of human LC3B. The exact sequence is proprietary.
Purification
Purified by antigen-affinity chromatography.
Conjugation
Unconjugated
RRID
AB_11176277
Note
For laboratory research use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
Purchasers shall not, and agree not to enable third parties to, analyze, copy, reverse engineer or otherwise attempt to determine the structure or sequence of the product.
TARGET
Synonyms
microtubule associated protein 1 light chain 3 beta , ATG8F , LC3B , MAP1A/1BLC3 , MAP1LC3B-a
Cellular Localization
Cytoplasm , Endomembrane system; Lipid-anchor , Cytoplasmic vesicle , autophagosome membrane; Lipid-anchor
Background
The product of this gene is a subunit of neuronal microtubule-associated MAP1A and MAP1B proteins, which are involved in microtubule assembly and important for neurogenesis. Studies on the rat homolog implicate a role for this gene in autophagy, a process that involves the bulk degradation of cytoplasmic component. [provided by RefSeq]
Database
Research Area
DATA IMAGES
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GTX127375 ICC/IF Image
LC3B antibody detects LC3B protein at autophagosome by immunofluorescent analysis.
Samples: HeLa cells mock (left) and treated with 50μM Chloroquine for 24 hr (right) were fixed in 4% paraformaldehyde at RT for 15 min.
Green: LC3B protein stained by LC3B antibody (GTX127375) diluted at 1:2000.
Red: alpha Tubulin, a cytoskeleton marker, stained by alpha Tubulin antibody [GT114] (GTX628802) diluted at 1:1000.
Blue: Hoechst 33342 staining.
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GTX127375 WB Image
Wild-type (WT) and LC3B knockout (KO) 293T cell extracts (30 μg) were separated by 15% SDS-PAGE, and the membrane was blotted with LC3B antibody (GTX127375) diluted at 1:500. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody, and the signal was developed with Trident ECL plus-Enhanced.
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GTX127375 WB Image
Untreated (–) and treated (+) HepG2 whole cell extracts (30 μg) were separated by 15% SDS-PAGE, and the membranes were blotted with LC3B antibody (GTX127375) diluted at 1:1000 and competitor's antibody (# Highly Cited Antibody ) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.
*The competitor is not affiliated with GeneTex and does not endorse this product.
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GTX127375 WB Image
LC3B antibody detects MAP1LC3B protein by western blot analysis.
A. 20 μg Huh7 whole cell lysate/extract (untreated)
B. 20 μg Huh7 whole cell lysate/extract (3uM-Thapsigargin treatment for 12hr)
LC3B antibody (GTX127375) dilution: 1:1500
The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.
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GTX127375 ICC/IF Image
LC3B antibody detects LC3B protein at autophagosome by immunofluorescent analysis.Sample: Mock and treated HeLa cells were fixed in 4% paraformaldehyde at RT for 15 min.Green: LC3B stained by LC3B antibody (GTX127375) diluted at 1:500.Red: phalloidin, a cytoskeleton marker, diluted at 1:200.Blue: Fluoroshield with DAPI (GTX30920).Scale bar= 10 μm.
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GTX127375 WB Image
HepG2 cells were untreated or treated with 3 μM thapsigargin for 16 and 24 hrs. Whole cell extracts (30 μg) were separated by 15% SDS-PAGE, and the membrane was blotted with LC3B antibody (GTX127375) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.
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GTX127375 WB Image
LC3B antibody detects LC3B protein in HCV-infected samples by western blot analysis.
A. 20 μg Huh7 whole cell lysate/extract (un-infected)
B. 20 μg Huh7 whole cell lysate/extract (HCV-infected)
LC3B antibody (GTX127375) dilution: 1:1500
The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.
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GTX127375 WB Image
LC3B antibody detects LC3B protein by western blot analysis. Various whole cell extracts (30 μg) were separated by 15% SDS-PAGE, and the membrane was blotted with LC3B antibody (GTX127375) diluted at a dilution of 1:1000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.
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GTX127375 IP Image
Immunoprecipitation of LC3B protein from U87-MG whole cell extracts using 5 μg of LC3B antibody (GTX127375).
Western blot analysis was performed using LC3B antibody (GTX127375).
EasyBlot anti-Rabbit IgG (GTX221666-01) was used as a secondary reagent.
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GTX127375 WB Image
LC3B antibody detects MAP1LC3B protein by western blot analysis.
A. 50 μg mouse brain lysate/extract
15% SDS-PAGE
LC3B antibody (GTX127375) dilution: 1:1000
The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.
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GTX127375 WB Image
Untreated (–) and treated (+) HeLa whole cell extracts (30 μg) were separated by 15% SDS-PAGE, and the membrane was blotted with LC3B antibody (GTX127375) diluted at 1:2500. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.
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GTX127375 WB Image
LC3B antibody detects MAP1LC3B protein by western blot analysis.
A. 50 μg Rat brain lysate/extract
15% SDS-PAGE
LC3B antibody (GTX127375) dilution: 1:1000
The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.
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GTX127375 WB Image
Untreated (–) and treated (+) HepG2 whole cell extract (30 μg) were separated by 15% SDS-PAGE, and the membrane was blotted with LC3B antibody (GTX127375) diluted at 1:500. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.
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GTX127375 ICC/IF Image
LC3B antibody detects LC3B protein at vesicle by immunofluorescent analysis.Sample: Mock and treated HepG2 cells were fixed in 4% paraformaldehyde at RT for 15 min.Green: LC3B stained by LC3B antibody (GTX127375) diluted at 1:500.Red: alpha Tubulin, a cytoskeleton marker, stained by alpha Tubulin antibody [GT114] (GTX628802) diluted at 1:1000.Blue: Fluoroshield with DAPI (GTX30920).
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GTX127375 FACS Image
LC3B antibody (GTX127375) detects LC3B protein by flow cytometry analysis.
Sample: HeLa cell fixed in 4% paraformaldehyde at 4oC for 5 min.
Brown: Unlabelled sample was also used as a control.
Blue: LC3B antibody (GTX127375) dilution: 1:100.
Acquisition of >20,000 events were collected using Argon ion laser (488nm) and 525/30 bandpass filter.
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GTX127375 IHC-P Image
LC3B antibody detects LC3B protein at cytoplasm by immunohistochemical analysis.Sample: Paraffin-embedded rat brain.LC3B stained by LC3B antibody (GTX127375) diluted at 1:500.Antigen Retrieval: Citrate buffer, pH 6.0, 15 min
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GTX127375 IHC-P Image
LC3B antibody detects LC3B protein at cytoplasm by immunohistochemical analysis.Sample: Paraffin-embedded rat cerebellum.LC3B stained by LC3B antibody (GTX127375) diluted at 1:500.Antigen Retrieval: Citrate buffer, pH 6.0, 15 min
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GTX127375 IHC-P Image
LC3B antibody detects LC3B protein at cytoplasm by immunohistochemical analysis.Sample: Paraffin-embedded mouse cerebellum.LC3B stained by LC3B antibody (GTX127375) diluted at 1:500.Antigen Retrieval: Citrate buffer, pH 6.0, 15 min
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GTX127375 WB Image
The data was published in the Cancer Med in 2020. PMID: 32324343
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GTX127375 WB Image
The data was published in the Cancer Med in 2020. PMID: 32324343
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GTX127375 WB Image
The data was published in the journal Sci Rep in 2017.PMID: 28195146
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GTX127375 WB Image
The data was published in the journal Sci Rep in 2017.PMID: 28195146
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GTX127375 WB Image
The data was published in the journal Biomolecules in 2019.PMID: 31505885
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GTX127375 WB Image
The data was published in the journal Antioxidants (Basel) in 2019.PMID: 31906147
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GTX127375 WB Image
The data was published in the journal Aging (Albany NY) in 2019. PMID: 31409760
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GTX127375 WB Image
The data was published in the journal Aging (Albany NY) in 2019. PMID: 31409760
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GTX127375 WB Image
The data was published in the journal Antioxidants (Basel) in 2019.PMID: 31906147
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GTX127375 WB Image
The data was published in the journal Antioxidants (Basel) in 2019.PMID: 31906147
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GTX127375 WB Image
The data was published in the 2019 in Int J Mol Sci. PMID: 30609861
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GTX127375 WB Image
The data was published in the Cancer Med in 2020. PMID: 32324343
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GTX127375 WB Image
The data was published in the Cancer Med in 2020. PMID: 32324343
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GTX127375 WB Image
The data was published in the journal Aging (Albany NY) in 2019. PMID: 31409760
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GTX127375 WB Image
The data was published in the journal Aging (Albany NY) in 2019. PMID: 31409760
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GTX127375 WB Image
The data was published in the journal Front Oncol in 2019. PMID: 31024851
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GTX127375 ICC/IF Image
The data was published in the journal Front Oncol in 2019. PMID: 31024851
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GTX127375 WB Image
The data was published in the Cancer Med in 2020. PMID: 32324343
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GTX127375 WB Image
The data was published in the journal Sci Rep in 2017.PMID: 28195146
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GTX127375 WB Image
The data was published in the journal Sci Rep in 2019.PMID: 31341250
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GTX127375 WB Image
The data was published in the journal Sci Rep in 2019. PMID: 30783186
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REFERENCE
REVIEW
LC3B antibody
Cat. No. GTX127375
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Rating
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( Average 4.4 based on 14 users reviews)
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Application
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Western Blot(WB)
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( Average 4.4 based on 13 users reviews)
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IHC (Paraffin sections)(IHC-P)
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( Average 5 based on 1 users reviews)
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Date :
Anonymous submitted on 19-Dec-2023
Application Tested :
WB
Sample Species :
Hu
Sample :
Starved HT29 and HCT116 cells with/without 10%FBS treatment
Amount used :
10μg
Blocking :
5% non-fat milk in TBST, 4°C, 1Hr
Primary Antibody :
1:1000, 4°C, 15Hr
Date :
Anonymous submitted on 08-Mar-2022
Application Tested :
IHC-P
Sample Species :
Rat
Sample :
Rat eye tissue section
Blocking :
5% goat serum, RT, 1Hr
Fixation :
PFA
Permeabilization /Antigen retrieval :
TBS-T / 1mM EDTA (pH8.0)
Primary Antibody :
1:100, 4°C, 12Hr
Date :
Anonymous submitted on 05-Aug-2021
Application Tested :
WB
Sample Species :
Hu
Sample :
U2OS cells
Amount used :
30 μg
Blocking :
5% non-fat dry milk in TBST, 25°C, 1Hr
Primary Antibody :
1:1000, 4°C, 12Hr
Figure Note :
0-30 μM cisplatin treatment for 24hr.
Date :
Anonymous submitted on 27-Jun-2018
Application Tested :
WB
Sample Species :
Hu
Amount used :
40 μg
Blocking :
5% BSA in TBST, 25°C, 1Hr
Primary Antibody :
1:1000, 4°C, 16Hr
Lane Description :
Lane 1: OEC-M1 cells, Lane 2: SAS cells
Date :
Anonymous submitted on 07-Mar-2017
Application Tested :
WB
Sample Species :
Ms
Sample :
Ovary tissue
Amount used :
20 μg
Blocking :
5% skim milk in TBST, 25°C, 1Hr
Primary Antibody :
1:1000, 4°C, 16Hr
Date :
Anonymous submitted on 06-May-2016
Application Tested :
WB
Sample Species :
Hu
Sample :
A549 cells
Amount used :
15μg
Blocking :
5% milk in TBST, 25°C, 1Hr
Primary Antibody :
1:2000, 4°C, 16Hr
Date :
Anonymous submitted on 25-Jun-2015
Application Tested :
WB
Sample Species :
Pig
Sample :
IPEC-J2 cells
Amount used :
5μg
Blocking :
5% non-fat milk, 22°C, 1Hr
Primary Antibody :
1:1000, 4°C, 14Hr
Lane Description :
Lane 1: IPEC-J2 cells.Lane 2: IPEC-J2 cells (10uM CCCP treatment for 12 hr).
Date :
Anonymous submitted on 24-Jun-2015
Application Tested :
WB
Sample Species :
Hu
Sample :
HaCaT cells
Amount used :
35μg
Blocking :
5% non-fat milk, 25°C, 1Hr
Primary Antibody :
1:1000, 4°C, 16Hr
Date :
Anonymous submitted on 23-Jan-2015
Application Tested :
WB
Sample Species :
Hu
Sample :
SK-N-SH cells
Amount used :
40μg
Blocking :
5% milk in TBST, 25°C, 1Hr
Primary Antibody :
1:1000, 4°C, 16Hr
Date :
Anonymous submitted on 15-Dec-2014
Application Tested :
WB
Sample Species :
Hu
Sample :
dendritic cells
Amount used :
20μg
Blocking :
10%milk in 0.05%TBST, 25°C, 1Hr
Primary Antibody :
1:1500, 4°C, 18Hr
Date :
Anonymous submitted on 26-May-2014
Application Tested :
WB
Sample Species :
Hu
Sample :
DU145 cells, PC3 cells
Amount used :
30μg
Blocking :
5% skim milk in PBST, 25°C, 0.5Hr
Primary Antibody :
1:1000, 25°C, 1Hr
Lane Description :
A:DU145 cells, B:PC3 cells
Date :
Anonymous submitted on 20-May-2013
Application Tested :
WB
Sample Species :
Rat
Sample :
H9C2 cells
Amount used :
10μg
Blocking :
5% skim milk, 25°C, 1Hr
Primary Antibody :
1:1000, 4°C, 16Hr
Date :
Anonymous submitted on 18-Mar-2013
Application Tested :
WB
Sample Species :
Hu
Sample :
A549 cells
Amount used :
20μg
Blocking :
5% milk in TBST, 25°C, 1Hr
Primary Antibody :
1:1000, 4°C, 16Hr
Date :
Anonymous submitted on 06-Feb-2013
Application Tested :
WB
Sample Species :
Ms
Sample :
3T3-L1 cells
Amount used :
40μg
Blocking :
5% skim milk, 37°C, 1Hr
Primary Antibody :
1:2500, 4°C, 16Hr
