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LC3B antibody

Anti-LC3B antibody used in Immunocytochemistry/ Immunofluorescence (ICC/IF). GTX127375
Anti-LC3B antibody used in Western Blot (WB). GTX127375
Anti-LC3B antibody used in Western Blot (WB). GTX127375
Anti-LC3B antibody used in Western Blot (WB). GTX127375
Anti-LC3B antibody used in Immunocytochemistry/ Immunofluorescence (ICC/IF). GTX127375
Anti-LC3B antibody used in Western Blot (WB). GTX127375
Anti-LC3B antibody used in Western Blot (WB). GTX127375
Anti-LC3B antibody used in Immunoprecipitation (IP). GTX127375
Anti-LC3B antibody used in Western Blot (WB). GTX127375
Anti-LC3B antibody used in Immunocytochemistry/ Immunofluorescence (ICC/IF). GTX127375
Anti-LC3B antibody used in Flow cytometry (FACS). GTX127375
Anti-LC3B antibody used in Western Blot (WB). GTX127375
Anti-LC3B antibody used in IHC (Paraffin sections) (IHC-P). GTX127375
Anti-LC3B antibody used in Western Blot (WB). GTX127375

Cat No. GTX127375

Host Rabbit
Clonality Polyclonal
Isotype IgG
Application WB, ICC/IF, IHC-P, FACS, IP
Reactivity Human, Mouse, Rat, Pig
Package
100 μl ($319),
25 μl ($169)
APPLICATION

Application Note

*Optimal dilutions/concentrations should be determined by the researcher.
Application Dilution
WB 1:500-1:3000
ICC/IF 1:100-1:1000
IHC-P 1:100-1:1000
FACS 1:50-1:200
IP 1:100-1:500
Not tested in other applications.

Calculated MW

15 kDa. ( Note )

Positive Control

NT2D1 , PC-3 , U87-MG , SK-N-SH , mouse brain , Rat brain , HepG2 (untreated) , HepG2 (3 μM Thapsigargin treatment for 12 hr) , HepG2 (3 μM Thapsigargin treatment for 16hr) , HepG2 (3 μM Thapsigargin treatment for 24hr) , Huh7 (un-infected) , Huh7 (HCV-infected) , HeLa , HeLa (50μM Chloroquine treatment for 24 hr)

Specificity/Sensitivity

Cross-reactivity may occur with other LC3 isoforms

Predict Reactivity

Zebrafish, Bovine, Japanese Medaka, Chimpanzee, Rhesus Monkey(>80% identity)
PROPERTIES

Form

Liquid

Buffer

1XPBS, 1% BSA, 20% Glycerol (pH7). 0.025% ProClin 300 was added as a preservative.

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

0.43 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

Note

For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
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
Anti-LC3B antibody used in Immunocytochemistry/ Immunofluorescence (ICC/IF). GTX127375

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.

Anti-LC3B antibody used in Western Blot (WB). GTX127375

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.

Anti-LC3B antibody used in Western Blot (WB). GTX127375

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.

Anti-LC3B antibody used in Western Blot (WB). GTX127375

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.

Anti-LC3B antibody used in Immunocytochemistry/ Immunofluorescence (ICC/IF). GTX127375

GTX127375 ICC/IF Image

LC3B antibody detects LC3B protein at autophagosome by immunofluorescent analysis.
Samples: Hep G2 cells mock (left) and treated with 3 μM Thapsigargin for 12 hrs (right) were fixed in ice-cold MeOH for 10 min and permeabilized with ice-cold acetone for 1 min.
Green: LC3B protein stained by LC3B antibody (GTX127375) diluted at 1:500.
Blue: Hoechst 33342 staining.
Scale bar = 10 μm.

Anti-LC3B antibody used in Western Blot (WB). GTX127375

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.

Anti-LC3B antibody used in Western Blot (WB). GTX127375

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.

Anti-LC3B antibody used in Immunoprecipitation (IP). GTX127375

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.

Anti-LC3B antibody used in Western Blot (WB). GTX127375

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.

Anti-LC3B antibody used in Immunocytochemistry/ Immunofluorescence (ICC/IF). GTX127375

GTX127375 ICC/IF Image

LC3B antibody detects LC3B protein at autophagosome by immunofluorescent analysis.
Sample: HeLa cells were fixed in 4% paraformaldehyde at RT for 15 min.
Green: LC3B protein stained by LC3B antibody (GTX127375) diluted at 1:200.
Red: phalloidin, a cytoskeleton marker, diluted at 1:50.
Blue: Hoechst 33342 staining.
Scale bar = 10 μm.

Anti-LC3B antibody used in Flow cytometry (FACS). GTX127375

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.

Anti-LC3B antibody used in Western Blot (WB). GTX127375

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.

Anti-LC3B antibody used in IHC (Paraffin sections) (IHC-P). GTX127375

GTX127375 IHC-P Image

LC3B antibody detects LC3B protein at cytoplasm in mouse brain by immunohistochemical analysis.
Sample: Paraffin-embedded mouse brain.
LC3B antibody (GTX127375) diluted at 1:500.

Antigen Retrieval: Citrate buffer, pH 6.0, 15 min

Anti-LC3B antibody used in Western Blot (WB). GTX127375

GTX127375 WB Image

Untreated (–) and treated (+) HepG2 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.

REFERENCE
REVIEW

LC3B antibody Cat No. GTX127375

Rating ( Average 4.3 based on 11 users reviews)
Application
Western Blot(WB) ( Average 4.3 based on 11 users reviews)
Date : Anonymous submitted on 27-Jun-2018
Score :
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
Score :
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
Score :
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
Score :
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
Score :
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
Score :
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
Score :
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
Score :
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
Score :
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
Score :
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
Score :
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
Application Reference
SDS
PBS.pdf
Glycerol.pdf
Proclin.pdf
Package List Price ($)
$ 319
$ 169