WB result of LC3B Recombinant Rabbit mAb
Primary antibody: LC3B Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: untreated HeLa whole cell lysate 20 µg
Lane 2: HeLa treated with 50 μM Chloroquine overnight whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 14 kDa
Observed MW: 14, 16 kDa
LC3B Recombinant Rabbit mAb (S-1599-142)
LC3B Recombinant Rabbit mAb (S-1599-142)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | LC3B |
| Synonyms | 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); MAP1ALC3; MAP1LC3B |
| Immunogen | Synthetic Peptide |
| Location | Cytoplasm, Cytoskeleton, Mitochondrion |
| Accession | Q9GZQ8 |
| Clone Number | S-1599-142 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC |
| Reactivity | Hu, Ms, Rt |
| Predicted Reactivity | Bv |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000 | Hu, Ms, Rt |
| IHC-P | 1:200-1:500 | Hu, Ms, Rt |
| ICC | 1:500 | Hu |
Background
LC3B protein is a crucial component in the autophagy process. It is a member of the LC3/GABARAP family of proteins. During autophagy, LC3B is cleaved by the enzyme Atg4 to generate the cytosolic form LC3B - I. Then LC3B - I is conjugated to phosphatidylethanolamine to form the membrane - bound form LC3B - II, which is recruited to the autophagosome membrane and plays an important role in the formation and maturation of autophagosomes. The conversion of LC3B - I to LC3B - II is often used as a marker to monitor autophagy activity. LC3B is involved in various cellular processes including the clearance of damaged organelles and protein aggregates, maintaining cellular homeostasis and responding to cellular stress such as nutrient deprivation and hypoxia. Dysregulation of LC3B - related autophagy has been implicated in the pathogenesis of many diseases like neurodegenerative disorders and cancer.
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Western Blot
WB result of LC3B Recombinant Rabbit mAb
Primary antibody: LC3B Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: untreated NIH/3T3 whole cell lysate 20 µg
Lane 2: NIH/3T3 treated with 50 μM Chloroquine overnight whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 14 kDa
Observed MW: 14, 15 kDa
WB result of LC3B Recombinant Rabbit mAb
Primary antibody: LC3B Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: untreated C6 whole cell lysate 20 µg
Lane 2: C6 treated with 50 μM Chloroquine overnight whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 14 kDa
Observed MW: 14, 15 kDa
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human cerebral cortex. Anti-LC3B antibody was used at 1/200 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human tonsil. Anti-LC3B antibody was used at 1/200 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human cervical squamous cell carcinoma. Anti-LC3B antibody was used at 1/200 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded mouse cerebral cortex. Anti-LC3B antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded rat cerebral cortex. Anti-LC3B antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Immunocytochemistry
ICC analysis of HeLa cells treated with Chloroquine (50μM,24h) (top panel) and untreated HeLa cells (below panel). Anti-LC3B antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
