Anti-LC3B Antibody, clone 12K5, ZooMAb® Rabbit Monoclonal
SIGMA/ZRB100 - recombinant, expressed in HEK 293 cells
Synonym: Autophagy-related protein LC3B; Autophagy-related ubiquitin-like modifier LC3B; MAP1 light chain 3-like protein 2; MAP1A/MAP1B LC3B; MAP1A/MAP1B light chain 3 B; Microtubule-
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) rat cerebral cortex (A) and human cerebral cortes (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:400 dilution of Cat. No. ZRB100, Anti-LC3B, clone 12K5 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit and DAB. Cytoplasmic/punctate cytoplasmic/membranous staining was observed in neurons of rat and human cerebral cortex.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) rat cerebral cortex (A) and human cerebral cortes (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:400 dilution of Cat. No. ZRB100, Anti-LC3B, clone 12K5 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit and DAB. Cytoplasmic/punctate cytoplasmic/membranous staining was observed in neurons of rat and human cerebral cortex.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) rat cerebral cortex (A) and human cerebral cortes (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:400 dilution of Cat. No. ZRB100, Anti-LC3B, clone 12K5 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit and DAB. Cytoplasmic/punctate cytoplasmic/membranous staining was observed in neurons of rat and human cerebral cortex.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HCT116 cells was performed using a 1:100 dilution of Cat. No. ZRB100, Anti-LC3B, clone 12K5 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoskeleton.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HCT116 cells was performed using a 1:100 dilution of Cat. No. ZRB100, Anti-LC3B, clone 12K5 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoskeleton.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HCT116 cells was performed using a 1:100 dilution of Cat. No. ZRB100, Anti-LC3B, clone 12K5 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoskeleton.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HCT116 cells treated with 50 μM Chloroquine overnight was performed using a 1:100 dilution of Cat. No. ZRB100, Anti-LC3B, clone 12K5 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoskeleton.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HCT116 cells treated with 50 μM Chloroquine overnight was performed using a 1:100 dilution of Cat. No. ZRB100, Anti-LC3B, clone 12K5 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoskeleton.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HCT116 cells treated with 50 μM Chloroquine overnight was performed using a 1:100 dilution of Cat. No. ZRB100, Anti-LC3B, clone 12K5 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoskeleton.
Immunofluorescence Enhanced Validation-Recombinant Antibody Technology Confocal analysis of human cerebral cortex tissue sections was performed using a 1:400 dilution of Cat. No. ZRB100, Anti-LC3B, clone 12K5 ZooMAb® Rabbit Monoclonal and visualized with a secondary antibody conjugated to Alexa Fluor® 488 (Green). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
Immunofluorescence Enhanced Validation-Recombinant Antibody Technology Confocal analysis of human cerebral cortex tissue sections was performed using a 1:400 dilution of Cat. No. ZRB100, Anti-LC3B, clone 12K5 ZooMAb® Rabbit Monoclonal and visualized with a secondary antibody conjugated to Alexa Fluor® 488 (Green). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
Immunofluorescence Enhanced Validation-Recombinant Antibody Technology Confocal analysis of human cerebral cortex tissue sections was performed using a 1:400 dilution of Cat. No. ZRB100, Anti-LC3B, clone 12K5 ZooMAb® Rabbit Monoclonal and visualized with a secondary antibody conjugated to Alexa Fluor® 488 (Green). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
Immunofluorescence Enhanced Validation-Recombinant Antibody Technology Confocal analysis of rat cerebral cortex tissue sections was performed using a 1:400 dilution of Cat. No. ZRB100, Anti-LC3B, clone 12K5 ZooMAb® Rabbit Monoclonal and visualized with a secondary antibody conjugated to Alexa Fluor® 488 (Green). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
Immunofluorescence Enhanced Validation-Recombinant Antibody Technology Confocal analysis of rat cerebral cortex tissue sections was performed using a 1:400 dilution of Cat. No. ZRB100, Anti-LC3B, clone 12K5 ZooMAb® Rabbit Monoclonal and visualized with a secondary antibody conjugated to Alexa Fluor® 488 (Green). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
Immunofluorescence Enhanced Validation-Recombinant Antibody Technology Confocal analysis of rat cerebral cortex tissue sections was performed using a 1:400 dilution of Cat. No. ZRB100, Anti-LC3B, clone 12K5 ZooMAb® Rabbit Monoclonal and visualized with a secondary antibody conjugated to Alexa Fluor® 488 (Green). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
Western Blotting Enhanced Validation-Recombinant Antibody Technology Lysates from untreated HeLa cells (Lane 1) and HeLa treated with Chloroquine (Lane 2) were probed with Cat. No. ZRB100, Anti-LC3B, clone 12K5 ZooMAb® Rabbit Monoclonal (1:1,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates LC3B (~15 kDa).
12 Principles of Green Chemistry: Principle 1—Waste Prevention. This product has a smaller environmental footprint through reduction of waste in the manufacturing process.
12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.
Chemical products should be designed to affect their desired function while minimizing their toxicity.
12 Principles of Green Chemistry: Principle 6—Design for Energy Efficiency. This product was re-engineered to utilize less energy in the manufacturing process.
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| biological source | rabbit (recombinant) |
| clone | 12K5, monoclonal |
| recombinant monoclonal | |
| conjugate | unconjugated |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| form | lyophilized |
| greener alternative category | Aligned |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry . |
| isotype | IgG |
| mol wt | calculated mol wt 14.69 kDa |
| observed mol wt ~15 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human, rat |
| storage temp. | 2-8°C |
| technique(s) | immunocytochemistry: suitable using 1:100 |
| immunofluorescence: 1:100-1:400 | |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): 1:100-1:400 | |
| western blot: suitable using 1:1,000 | |
| UniProt accession no. | Q9GZQ8  |
| Application: | Anti-LC3B, clone 12K5 ZooMAb, Cat. No. ZRB100, is a recombinant Rabbit Monoclonal Antibody that detects Microtubule-associated proteins 1A/1B light chain 3B and has been tested for use in Immunocytochemistry, Immunofluorescence,Immuno |
| Application: | Immunohistochemistry (Paraffin) Analysis: A 1:400 dilution from a representative lot detected LC3B in rat brain and human brain tissue sections. Immunofluorescence Analysis: A 1:400 dilution from a representative lot detected LC3B in human brain and rat brain tissue sections. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected LC3B in HCT116 cells treated with 50 uM Chloroquine overnight. |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. |
| General description: | We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information. |
| General description: | ZooMAb antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb antibodies are reliably available and ready to ship when you need them. Learn more about ZooMAb here. |
| Immunogen: | KLH-conjugated linear peptide corresponding to 14 amino acids from the N-terminal region of human Microtubule-associated proteins 1A/1B light chain 3B (LC3B). |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS with 5% Trehalose, normal appearance a coarse or translucent resin. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL. |
| Reconstitution: | 120 μg/mL after reconstitution at 25 μL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 12K5 is a ZooMAb rabbit recombinant monoclonal antibody that detects Microtubule-associated proteins 1A/1B light chain 3B (LC3B) in human and rat. It targets an epitope with in 14 amino acids from the N-terminal region. |
| Storage and Stability: | Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws. |
| Target description: | Microtubule-associated proteins 1A/1B light chain 3B (UniProt: Q9GZQ8; also known as Autophagy-related protein LC3B, Autophagy-related ubiquitin-like modifier LC3B, MAP1 light chain 3-like protein 2, MAP1A/MAP1B light chain 3 B, MAP1A/MAP1B LC3B, Microtubule-associated protein 1 light chain 3 beta) is encoded by the MAP1LC3B (also known as MAP1ALC3) gene (Gene ID: 81631) in human. LC3 is ubiquitously distributed in all eukaryotes and is expressed as three splice variants known as LC3A, LC3B, and LC3C. They possess a ubiquitin-like structure despite differences in their respective amino acid sequences. The amino-terminus of LC3 is composed of an alpha-helix that varies among the 3 members. However, its C-terminus is conserved and consists of a ubiquitin core composed of beta strands with hydrophobic pockets. The ubiquitin core region is responsible for its interaction with Atg core machinery proteins. LC3 is cleaved by Atg4 between amino acids 120-121 to form LC3-I and expose its C-terminal glycine. LC3-I then conjugates to phosphatidylethanolamine (PE) by Atg7, Atg3, and the Atg12 complex to generate LC3-II, which is inserted into autophagosome membrane. LC3B is reported to be most abundant in heart, brain, skeletal muscle and testis and only a trivial expression is observed in the liver. Higher expression of LC3B in triple negative breast cancer is associated with poor outcomes in patients. It plays a role in mitophagy and regulates mitochondrial quantity and quality by eliminating the mitochondria to a basal level to fulfill cellular energy requirements and preventing excess reactive oxygen species production. C3B is synthesized with a propeptide (aa 121-125), which is cleaved off to produce the mature form. LC3B can undergo phosphorylation at threonine 12 by protein kinase A (PKA) and this phosphorylation inhibits its conjugation to phosphatidylethanolamine. This ZooMAb recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Lee, YK and Lee JA (2016). BMB Rep. 49(8): 424-430). |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |