Anti-LC3B antibody - N-terminal (ab229327)
Key features and details
- Rabbit polyclonal to LC3B - N-terminal
- Suitable for: WB, Flow Cyt, ICC/IF, IP, IHC-P
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
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- Research with confidence – consistent and reproducible results with every batch
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Overview
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Product name
Anti-LC3B antibody - N-terminal
See all LC3B primary antibodies -
Description
Rabbit polyclonal to LC3B - N-terminal -
Host species
Rabbit -
Tested applications
Suitable for: WB, Flow Cyt, ICC/IF, IP, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Cow, Chimpanzee, Zebrafish, Rhesus monkey, Medaka fish
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Immunogen
Synthetic peptide within Human LC3B (N terminal). The exact sequence is proprietary. (Carrier-protein conjugated).
Database link: Q9GZQ8 -
Positive control
- ICC/IF: HeLa cells untreated and treated with 50µM Chloroquine for 24 hr; Hep G2 cells untreated and treated with 3 µM Thapsigargin for 12 hrs. WB: Huh7 whole cell lysate; NTERA-2 cl.D1 [NT2/D1], PC-3, U-87 MG and SK-N-SH whole cell extracts; Mouse and rat brain lysates; Hep G2 cell lysate untreated and treated with 3 µM Thapsigargin; HeLa whole cell extract untreated and treated with 50µM Chloroquine for 24 hr. IHC-P: Mouse brain tissue. IP: U-87 MG whole cell extract. Flow Cyt: HeLa cells.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.00
Preservative: 0.025% Proclin 300
Constituents: 78% PBS, 1% BSA, 20% Glycerol (glycerin, glycerine) -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab229327 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB |
1/500 - 1/3000. Predicted molecular weight: 15 kDa.
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| Flow Cyt |
1/50 - 1/200.
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| ICC/IF |
1/100 - 1/1000.
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| IP |
1/100 - 1/500.
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| IHC-P |
1/100 - 1/1000.
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| Notes |
|---|
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WB
1/500 - 1/3000. Predicted molecular weight: 15 kDa. |
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Flow Cyt
1/50 - 1/200. |
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ICC/IF
1/100 - 1/1000. |
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IP
1/100 - 1/500. |
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IHC-P
1/100 - 1/1000. |
Target
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Function
Probably involved in formation of autophagosomal vacuoles (autophagosomes). -
Tissue specificity
Most abundant in heart, brain, skeletal muscle and testis. Little expression observed in liver. -
Sequence similarities
Belongs to the MAP1 LC3 family. -
Post-translational
modificationsThe precursor molecule is cleaved by APG4B/ATG4B to form LC3-I. This is activated by APG7L/ATG7, transferred to ATG3 and conjugated to phospholipid to form LC3-II. -
Cellular localization
Cytoplasm > cytoskeleton. Endomembrane system. Cytoplasmic vesicle > autophagosome membrane. LC3-II binds to the autophagic membranes. - Information by UniProt
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Database links
- Entrez Gene: 746157 Chimpanzee
- Entrez Gene: 408001 Cow
- Entrez Gene: 81631 Human
- Entrez Gene: 67443 Mouse
- Entrez Gene: 64862 Rat
- Entrez Gene: 695419 Rhesus monkey
- Entrez Gene: 322425 Zebrafish
- Omim: 609604 Human
see all -
Alternative names
- ATG8F antibody
- Autophagy-related protein LC3 B antibody
- Autophagy-related ubiquitin-like modifier LC3 B antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LC3B antibody - N-terminal (ab229327)
Paraffin-embedded mouse brain tissue stained for LC3B using ab229327 at 1/500 dilution in immunohistochemical analysis.
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Western blot - Anti-LC3B antibody - N-terminal (ab229327)All lanes : Anti-LC3B antibody - N-terminal (ab229327) at 1/1500 dilution
Lane 1 : Huh7 whole cell lysate (untreated)
Lane 2 : Huh7 whole cell lysate (3 µM Thapsigargin treatment for 12 hr)
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP-conjugated anti-rabbit IgG
Predicted band size: 15 kDa -
Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody - N-terminal (ab229327)4% paraformaldehyde-fixed HeLa (human epithelial cell line from cervix adenocarcinoma) cells, mock treated (left panel) or treated with 50 μM Chloroquine for 24 hr (right panel), stained for LC3B (green) using ab229327 at 1/2000 dilution in ICC/IF.
Red: alpha Tubulin, a cytoskeleton marker, stained by alpha Tubulin antibody at 1/1000 dilution.
Blue: Hoechst 33342 staining.
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Western blot - Anti-LC3B antibody - N-terminal (ab229327)All lanes : Anti-LC3B antibody - N-terminal (ab229327) at 1/1500 dilution
Lane 1 : Huh7 whole cell lysate (un-infected)
Lane 2 : Huh7 whole cell lysate (HCV-infected)
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP-conjugated anti-rabbit IgG
Predicted band size: 15 kDa -
Western blot - Anti-LC3B antibody - N-terminal (ab229327)All lanes : Anti-LC3B antibody - N-terminal (ab229327) at 1/1000 dilution
Lane 1 : NTERA-2 cl.D1 [NT2/D1] (human malignant pluripotent embryonic carcinoma cell line) whole cell extract
Lane 2 : PC-3 (human prostate adenocarcinoma cell line) whole cell extract
Lane 3 : U-87 MG (human glioblastoma-astrocytoma epithelial cell line) whole cell extract
Lane 4 : SK-N-SH (human neuroblastoma cell line) whole cell extract
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : HRP-conjugated anti-rabbit IgG
Predicted band size: 15 kDa15% SDS-PAGE gel.
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Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody - N-terminal (ab229327)Ice-cold methanol-fixed, ice-cold acetone permeabilized HepG2 (human liver hepatocellular carcinoma cell line) cells, mock treated (left panels) or treated with with 3 μM Thapsigargin for 12 hrs (right panels), stained for LC3B (green) using ab229327 at 1/500 dilution in ICC/IF.
Blue: Hoechst 33342 staining.
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Immunoprecipitation - Anti-LC3B antibody - N-terminal (ab229327)LC3B was immunoprecipitated from U-87 MG (human glioblastoma-astrocytoma epithelial cell line) whole cell extract (1 mg for IP, 20% of IP loaded) with 5 µg ab229327. Western blot was performed from the immunoprecipitate using ab229327. Anti-Rabbit IgG was used as a secondary reagent.
Lane 1: U-87 MG whole cell extract.
Lane 2: Control IgG IP in U-87 MG whole cell extract.
Lane 3: ab229327 IP in U-87 MG whole cell extract.
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Flow Cytometry - Anti-LC3B antibody - N-terminal (ab229327)Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling LC3B with ab229327 at 1/100 dilution (blue) compared with an unlabelled sample (brown).
Acquisition of >20,000 events were collected using Argon ion laser (488nm) and 525/30 bandpass filter.
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Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody - N-terminal (ab229327)4% paraformaldehyde-fixed HeLa (human epithelial cell line from cervix adenocarcinoma) cells, mock treated (left panels) or treated with 50 μM Chloroquine for 24 hr (right panels), stained for LC3B (green) using ab229327 at 1/200 dilution in ICC/IF.
Red: phalloidin, a cytoskeleton marker, at 1/50 dilution.
Blue: Hoechst 33342 staining.
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Western blot - Anti-LC3B antibody - N-terminal (ab229327)Anti-LC3B antibody - N-terminal (ab229327) at 1/1000 dilution + Mouse brain lysate at 50 µg
Secondary
HRP-conjugated anti-rabbit IgG
Predicted band size: 15 kDa15% SDS-PAGE gel.
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Western blot - Anti-LC3B antibody - N-terminal (ab229327)Anti-LC3B antibody - N-terminal (ab229327) at 1/1000 dilution + Rat brain lysate at 50 µg
Secondary
HRP-conjugated anti-rabbit IgG
Predicted band size: 15 kDa15% SDS-PAGE gel.
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Western blot - Anti-LC3B antibody - N-terminal (ab229327)All lanes : Anti-LC3B antibody - N-terminal (ab229327) at 1/1000 dilution
Lane 1 : Untreated HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 2 : HepG2 whole cell lysate treated with 3 µM thapsigargin for 12 hours
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : HRP-conjugated anti-rabbit IgG
Predicted band size: 15 kDaLower panel: Beta-actin antibody at 1/20000 dilution.
15% SDS-PAGE gel.
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Western blot - Anti-LC3B antibody - N-terminal (ab229327)All lanes : Anti-LC3B antibody - N-terminal (ab229327) at 1/1000 dilution
Lane 1 : Untreated HepG2 (human liver hepatocellular carcinoma cell line) whole cell extract
Lane 2 : HepG2 whole cell extract treated with 3 µM thapsigargin for 16 hours
Lane 3 : HepG2 whole cell extract treated with 3 µM thapsigargin for 24 hours
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : HRP-conjugated anti-rabbit IgG
Predicted band size: 15 kDa15% SDS-PAGE gel.
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Western blot - Anti-LC3B antibody - N-terminal (ab229327)All lanes : Anti-LC3B antibody - N-terminal (ab229327) at 1/2500 dilution
Lane 1 : Untreated HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell extract
Lane 2 : HeLa whole cell extract treated with 50µM Chloroquine for 24 hours
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : HRP-conjugated anti-rabbit IgG
Predicted band size: 15 kDa15% SDS-PAGE gel.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (10)
ab229327 has been referenced in 10 publications.
- Qiu S et al. Hepatic lipid accumulation induced by a high-fat diet is regulated by Nrf2 through multiple pathways. FASEB J 36:e22280 (2022). PubMed: 35394671
- Li L et al. Effect of Platycodon grandiflorus Polysaccharide on M1 Polarization Induced by Autophagy Degradation of SOCS1/2 Proteins in 3D4/21 Cells. Front Immunol 13:934084 (2022). PubMed: 35844489
- Saleh SR et al. The Synergetic Effect of Egyptian Portulaca oleracea L. (Purslane) and Cichorium intybus L. (Chicory) Extracts against Glucocorticoid-Induced Testicular Toxicity in Rats through Attenuation of Oxidative Reactions and Autophagy. Antioxidants (Basel) 11:N/A (2022). PubMed: 35883763
- Dou X et al. Thymol Alleviates LPS-Induced Liver Inflammation and Apoptosis by Inhibiting NLRP3 Inflammasome Activation and the AMPK-mTOR-Autophagy Pathway. Nutrients 14:N/A (2022). PubMed: 35889766
- Wang L et al. Danlou Tablet Activates Autophagy of Vascular Adventitial Fibroblasts Through PI3K/Akt/mTOR to Protect Cells From Damage Caused by Atherosclerosis. Front Pharmacol 12:730525 (2021). PubMed: 34867337
- Meng CY et al. MicroRNA-22 mediates the cisplatin resistance of osteosarcoma cells by inhibiting autophagy via the PI3K/Akt/mTOR pathway. Oncol Rep 43:1169-1186 (2020). PubMed: 32323781
- Hao J et al. Platycodon grandifloras polysaccharides inhibit mitophagy injury induced by Cr (VI) in DF-1 cells. Ecotoxicol Environ Saf 202:110901 (2020). PubMed: 32593805
- Meng CY et al. MicroRNA-22 regulates autophagy and apoptosis in cisplatin resistance of osteosarcoma. Mol Med Rep 22:3911-3921 (2020). PubMed: 33000186
- Li L et al. Puerarin 6?-O-xyloside suppressed HCC via regulating proliferation, stemness, and apoptosis with inhibited PI3K/AKT/mTOR. Cancer Med 9:6399-6410 (2020). PubMed: 32691991
- Liu K et al. Protective Effect of Purple Tomato Anthocyanidin on Chromium(VI)-Induced Autophagy in LMH Cells by Inhibiting Endoplasmic Reticulum Stress. Biol Trace Elem Res N/A:N/A (2019). PubMed: 31264128