Name: Anti-LC3B antibody [EPR18709] - Autophagosome Marker
SKU: ab192890
Rating: 5 (19 reviews)

Anti-LC3B antibody [EPR18709] - Autophagosome Marker is a rabbit recombinant monoclonal antibody that is used to detect LC3B in ICC/IF, IHC-P, Western blot. Suitable for Human, Mouse, Rat samples.

- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Specificity confirmed with MAP1LC3B knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Antibody clone EPR18709 is cited in over 700 publications

Images

Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (AB192890), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IHC-P	IP	WB	ICC/IF
Human	Tested	Not recommended	Tested	Tested
Mouse	Expected	Not recommended	Tested	Expected
Rat	Expected	Not recommended	Tested	Expected

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 0.1-1 µg/mL	Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/2000	Notes -
Species Rat	Dilution info 1/2000	Notes -
Species Human	Dilution info 1/2000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1 µg/mL	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Associated Products

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Target data

See full target information MAP1LC3B

Function

Ubiquitin-like modifier involved in formation of autophagosomal vacuoles (autophagosomes) (PubMed:20418806, PubMed:23209295, PubMed:28017329). Plays a role in mitophagy which contributes to regulate mitochondrial quantity and quality by eliminating the mitochondria to a basal level to fulfill cellular energy requirements and preventing excess ROS production (PubMed:23209295, PubMed:28017329). In response to cellular stress and upon mitochondria fission, binds C-18 ceramides and anchors autophagolysosomes to outer mitochondrial membranes to eliminate damaged mitochondria (PubMed:22922758). While LC3s are involved in elongation of the phagophore membrane, the GABARAP/GATE-16 subfamily is essential for a later stage in autophagosome maturation (PubMed:20418806, PubMed:23209295, PubMed:28017329). Promotes primary ciliogenesis by removing OFD1 from centriolar satellites via the autophagic pathway (PubMed:24089205). Through its interaction with the reticulophagy receptor TEX264, participates in the remodeling of subdomains of the endoplasmic reticulum into autophagosomes upon nutrient stress, which then fuse with lysosomes for endoplasmic reticulum turnover (PubMed:31006537, PubMed:31006538). Upon nutrient stress, directly recruits cofactor JMY to the phagophore membrane surfaces and promotes JMY's actin nucleation activity and autophagosome biogenesis during autophagy (PubMed:30420355).

Alternative names

MAP1ALC3, MAP1LC3B, Microtubule-associated proteins 1A/1B light chain 3B, Autophagy-related protein LC3 B, Autophagy-related ubiquitin-like modifier LC3 B, MAP1 light chain 3-like protein 2, MAP1A/MAP1B light chain 3 B, Microtubule-associated protein 1 light chain 3 beta, MAP1A/MAP1B LC3 B

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR18709
Purification technique: Affinity purification Protein A
Specificity: FURTHER INFORMATION ON SPECIFICITY (Chinese Version) available under the support & downloads section.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in ICC/IF, IHC-P and WB.

Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890) was first used in a scientific publication in 2017 and has been cited over 468 times in peer reviewed journals. It's performance in Western Blot in human, mouse and rat samples is trusted by the scientific community.

Abcam's high quality manufacturing and validation processes ensure Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.

The specificity of Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890) has been confirmed by Western Blot testing in LC3B knockout HepG2 cells (ab277828).

Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890) has 17 independent reviews from customers.

Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890) specifically detects LC3B (UniProt ID: Q9GZQ8; Molecular weight: 15kDa) and is sold in a convenient trial size to enable initial testing (20 µL) and larger sizes for subsequent scaling up experiments (100 µL and 1 mL).

Conjugation-ready, carrier free format available for antibody clone EPR18709 - Anti-LC3B antibody [EPR18709] - BSA and Azide free ab221794.

Antibody clone EPR18709 is also available pre-conjugated to a variety of labels for your convenience - Alexa Fluor^® 488, Alexa Fluor^® 647, Alexa Fluor^® 555 (Alexa Fluor® 488 Anti-LC3B antibody [EPR18709] - Autophagosome Marker ab225382, Alexa Fluor® 647 Anti-LC3B antibody [EPR18709] - Autophagosome Marker ab225383, Alexa Fluor® 555 Anti-LC3B - Autophagosome Marker antibody [EPR18709] ab307768).

LC3B is a protein involved in autophagy, a process that helps maintain cellular homeostasis by degrading damaged components. In oncology, LC3B is used as a marker to assess autophagy levels in cancer cells, which can influence tumor progression and response to therapy.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

LC3B also known as Microtubule-associated protein 1A/1B-light chain 3 is a core protein in autophagy processes. LC3B protein has a molecular weight of approximately 14 kDa. Researchers often utilize LC3B as a marker to study autophagic activity especially through LC3B immunofluorescence and LC3B Western blot analyses. This protein is predominantly found in the cytoplasm and is widely expressed in various tissues with higher expression in tissues undergoing high turnover or stress.

Biological function summary

LC3B contributes to the formation of autophagosomes playing an essential role in autophagy. During autophagosome formation LC3B conjugates with phosphatidylethanolamine which localizes it to autophagosome membranes. This lipidated form is important for the elongation and closure of autophagosomes. LC3B is also a component of the autophagy-related protein complex collaborating with other Atg proteins to ensure the proper degradation of cytoplasmic contents.

Pathways

LC3B plays an important role in the autophagy pathway important for cellular homeostasis and stress response. This pathway interfaces with the mTOR signaling pathway where LC3B helps regulate mTOR activity by controlling autophagosome dynamics. LC3B is associated with proteins like Beclin-1 and p62/SQSTM1 within these pathways coordinating the sequestration and degradation of proteins and organelles.

Associated diseases and disorders

LC3B is implicated in cancer and neurodegenerative diseases. Altered LC3B levels and function are observed in cancers linking this protein to tumorigenesis through its role in autophagy. In neurodegenerative disorders like Parkinson's disease LC3B interacts with proteins such as alpha-synuclein influencing the accumulation of protein aggregates and neuronal health. The modulation of LC3B activity is of interest in therapeutic strategies targeting these diseases.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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12 product images

Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
Lane 1: Wild-type HAP1 cell lysate (20 μg)
Lane 2: LC3B knockout HAP1 cell lysate (20 μg)
Lane 3: Human brain tissue lysate (20 μg)
Lane 4: U-87 MG cell lysate (20 μg)
Lanes 1 - 4: Merged signal (red and green).
Green -target observed at 14 and 16 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
This western blot image is a comparison between ab192890 and a competitor's top cited rabbit polyclonal antibody.
All lanes: Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
Predicted band size: 15 kDa
Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
LC3B Immunocytochemistry/ Immunofluorescence staining using rabbit Anti-LC3B antibody
ab192890 staining LC3B in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells +/- Chloroquine (50μM 24 hours).
The cells were fixed with 100% methanol (5 min) permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab192890 at 1μg/ml and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 594) at 1/250 dilution (shown in pseudocolor red) followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
Alexa Fluor^® 488 (Alexa Fluor® 647 Anti-LC3B antibody [EPR18709] - Autophagosome Marker ab225383) and Alexa Fluor^® 647 (Alexa Fluor® 488 Anti-LC3B antibody [EPR18709] - Autophagosome Marker ab225382) conjugated versions are available for this clone.
Immunoprecipitation - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
LC3B was immunoprecipitated from 1 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab192890 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab192890 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10,000 dilution.
Lane 1: HeLa whole cell lysate 10 μg (Input).
Lane 2: ab192890 IP in HeLa whole cell lysate.
Lane 3: Rabbit IgG,monoclonal [EPR25A] Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab192890 in HeLa whole cell cell lysate.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 30 seconds.
All lanes: Immunoprecipitation - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
Predicted band size: 15 kDa
Exposure time: 30s
Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
LC3B Immunocytochemistry/ Immunofluorescence staining using rabbit Anti-LC3B antibody
ab192890 staining LC3B in HAP1 cells (wildtype and MAP1LC3B knockout) +/- Chloroquine (50μM 24 hours).
The cells were fixed with 100% methanol (5 min) permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab192890 at 1 μg/ml and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 594) at 1/250 dilution (shown in pseudocolor red) followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
IHC image of LC3B staining in a section of formalin-fixed paraffin-embedded normal human cerebral cortex* performed on a Leica BOND^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with Anti-LC3B antibody [EPR18709] - BSA and Azide free ab221794, 0.1ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
This data was developed using the same antibody clone in a different buffer formulation containing only PBS (Anti-LC3B antibody [EPR18709] - BSA and Azide free ab221794).
Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
Lane 1: Wild-type HAP1 cell lysate (20 μg)
Lane 2: LC3B knockout HAP1 cell lysate (20 μg)
Lane 3: Human brain tissue lysate (20 μg)
Lane 4: U-87 MG cell lysate (20 μg)
Lanes 1 - 4: Merged signal (red and green). Green - ab192890 observed at 14 and 16 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.

ab192890 was shown to specifically react with LC3B in wild-type HAP1 cells. No band was observed when LC3B knockout samples were examined. Wild-type and LC3B knockout samples were subjected to SDS-PAGE. ab192890 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (loading control to GAPDH) were diluted 1/2000 and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
Predicted band size: 15 kDa
Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure times: Lane 1-6: 3 minutes; Lane 7 and 8: 30 seconds.
All lanes: Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890) at 1/2000 dilution
Lane 1: Human brain lysate at 20 µg
Lane 2: U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate at 20 µg
Lane 3: C6 (Rat glial tumor cell line) whole cell lysate at 20 µg
Lane 4: RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 20 µg
Lane 5: Mouse heart lysate at 20 µg
Lane 6: Rat heart lysate at 20 µg
Lane 7: Mouse brain lysate at 20 µg
Lane 8: Rat brain lysate at 20 µg
Secondary
All lanes: Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 15 kDa
Observed band size: 14 kDa, 16 kDa
Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
Different batches of ab192890 were tested onU-87 MG (Human glioblastoma-astrocytoma epithelial cell line) lysate at 0.9 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 14,16 kDa.
All lanes: Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
Predicted band size: 15 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
ab192890 staining LC3B in paraffin embedded human astrocytoma tissue by Immunohistochemistry. Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins. Samples were incubated with primary antibody at 1/1000 dilution for 30 mins at room temperature. Ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used as the secondary antibody. Hematoxylin was used as a counterstain. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
LC3B Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of human cortex using rabbit Anti-LC3B antibody
Immunohistochemical analysis of formalin fixed paraffin embedded human cortex labelling LC3B with ab192890 at a dilution of 0.1 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32min with ULTRA cell conditioning solution (CC1 pH8.5) . ab192890 anti LC3B antibody was incubated at 37°C for 16min. Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control
Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
False colour image of Western blot: Anti-LC3B antibody [EPR18709] - Autophagosome Marker staining at 1/2000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab192890 was shown to bind specifically to LC3B. A band was observed at 16/14 kDa (yellow arrows) in treated wild-type HepG2 cell lysates with no signal observed at this size in MAP1LC3B knockout cell line ab277828 (knockout cell lysate ab283796). To generate this image, wild-type and MAP1LC3B knockout HepG2 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890) at 1/1000 dilution
Lane 1: Wild-type HepG2 untreated control cell lysate at 20 µg
Lane 2: Wild-type HepG2 Treated Chloroquine (50 uM, 16 h) cell lysate at 20 µg
Lane 2: Western blot - Human MAP1LC3B knockout Hep G2 cell line (ab277828)
Lane 2: Western blot - Human MAP1LC3B knockout Hep G2 cell lysate (ab283796)
Lane 3: MAP1LC3B knockout HepG2 untreated control cell lysate at 20 µg
Lane 4: MAP1LC3B knockout HepG2 Treated Chloroquine (50 uM, 16 h) cell lysate at 20 µg
Lane 5: U-87 MG cell lysate at 20 µg
Lane 6: PC-12 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 15 kDa
Observed band size: 14 kDa, 16 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
Tissue Microarrays stained for Anti-LC3B antibody [EPR18709] - Autophagosome Marker using ab192890 in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negaive (cross mark) staining per sample type tested.

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

The section was incubated with ab192890 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.