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AB322912

Anti-LC3B antibody [EPR18709] - Autophagosome Marker – Chicken IgY (Chimeric)

  • 20ul selling size
  • BOND RX™ Validated
  • Recombinant
  • KO Validated
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Chicken recombinant monoclonal LC3B EPR18709 antibody. Suitable for IHC-P and ICC/IF. Reacts with human and mouse samples.

This antibody is the chimeric Chicken version of clone EPR18709, available in rabbit as ab192890 with over 750 publications. Same clone, same staining pattern, different species antibody backbone.
Learn more about chimeric antibodies

View Alternative Names

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

4 Images
Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody [EPR18709] - Autophagosome Marker – Chicken IgY (Chimeric) (AB322912)
  • ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody [EPR18709] - Autophagosome Marker – Chicken IgY (Chimeric) (AB322912)

ab322912 staining LC3B in HAP1 cells (wildtype and MAP1LC3B knockout) +/- Chloroquine (50µM 24 hours) cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-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 ab322912 at 1µg/ml (shown in green) and ab202272, Alexa Fluor® 594 Rabbit monoclonal [EP1332Y] to alpha Tubulin - Microtubule Marker (shown in pseudocolour magenta). Cells were then incubated with ab150173, Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LC3B antibody [EPR18709] - Autophagosome Marker – Chicken IgY (Chimeric) (AB322912)
  • IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LC3B antibody [EPR18709] - Autophagosome Marker – Chicken IgY (Chimeric) (AB322912)

Immunofluorescence staining of LC3B in sections of formalin-fixed paraffin-embedded human brain (positive) and human heart muscle (negative)*. Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with ab322912 at 1/100 dilution and then incubated for 1 hour with ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LC3B antibody [EPR18709] - Autophagosome Marker – Chicken IgY (Chimeric) (AB322912)
  • IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LC3B antibody [EPR18709] - Autophagosome Marker – Chicken IgY (Chimeric) (AB322912)

Immunofluorescence staining of LC3B in sections of formalin-fixed paraffin-embedded mouse brain (positive) and mouse liver (negative). Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with ab322912 at 1/100 dilution and then incubated for 1 hour with ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody [EPR18709] - Autophagosome Marker – Chicken IgY (Chimeric) (AB322912)
  • ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody [EPR18709] - Autophagosome Marker – Chicken IgY (Chimeric) (AB322912)

ab322912 staining LC3B in NIH/3T3 +/- Chloroquine (50µM 24 hours) cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-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 ab322912 at 1µg/ml (shown in green) and ab202272, Alexa Fluor® 594 Rabbit monoclonal [EP1332Y] to alpha Tubulin - Microtubule Marker (shown in pseudocolour magenta). Cells were then incubated with ab150173, Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

Key facts

Host species

Chicken

Clonality

Monoclonal

Clone number

EPR18709

Isotype

IgY

Carrier free

No

Reacts with

Human, Mouse

Applications

ICC/IF, IHC-P

applications

Immunogen

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

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/500", "ICCIF-species-notes": "<p></p>", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/100", "IHCP-species-notes": "<p></p> Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol." }, "Mouse": { "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/500", "ICCIF-species-notes": "<p></p>", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/100", "IHCP-species-notes": "<p></p> Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol." } } }

Product details

This chicken monoclonal chimeric antibody has been engineered from a RabMAb parent antibody (ab192890). By necessity, some rabbit sequence is retained as part of the variable domain. When multiplexing with other rabbit-derived antibodies, using cross absorbed Fc-reactive secondary antibodies are recommended.

What are the advantages of a recombinant monoclonal antibody?
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.

Properties and storage information

Form
Liquid
Purification technique
Affinity purification Thiophilic Resin
Storage buffer
pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
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

Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Target data

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).
See full target information MAP1LC3B

Product promise

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