WB
Western blot analysis using LC3A/B antibody against B16(1),A549(2),Nih3t3(3) cell . 12% SDS-PAGE gel.Sample loading: 20μg /lane. Transfer the proteins onto a PVDF membrane (OM790003), and block it with TBST (OM750016) plus skimmed milk powder(OM750084) for one hour. Dilute the primary antibody with the antibody diluent (OM750012) at a ratio of 1:1000, and incubate it overnight at 4°C. Wash the membrane three times with TBST (OM750016), 5 minutes each time. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP (OM643487), at a ratio of 1:20000 and incubate for one hour. Wash the membrane three times with TBST (OM750016) again, 5 minutes each time. Use ECL (OM625701) for luminescence.staining time: 60S.ELISA
Microtiter plate wells were coated with 100 ng/well coating antigen and incubated with serial dilutions of the antibody. Following HRP-conjugated secondary antibody and substrate development, OD450 was measured. The absorbance decreased progressively with increasing antibody dilution, yielding a sigmoidal titration curve that demonstrates the high antigen-binding activity of the polyclonal antibody.ICC/IF
Immunofluorescence analysis of NIH3T3 cells using LC3A/B antibody (green). Blue: DAPI fluorescent DNA dye. Red: Actin filaments have been labeled with Omnimabs® 594-Phalloidin.Cells are fixed in 4% paraformaldehyde at room temperature for 15 minutes. Then, they are permeabilized with a PBS (OM750003) solution containing 0.1% Triton X-100(OM750021) at room temperature for 15 minutes. Subsequently, the cells are blocked with 10% non - immune goat serum(OM760028) at room temperature for 1 hour.The cells are incubated overnight at 4°C with the primary antibody diluted 1:100 in PBS. The secondary antibody, Omnimabs® 488 Goat Ant-Rabbit IgG(H&L) (Green,OM643486), is diluted at a ratio of 1:400 and incubated with the cells for 1 hour.Nuclear DNA is labeled with DAPI (Blue,OM643160). F-actin is stained with Omnimabs® 594-Phalloidin (Red,OM750007) diluted 1:100 for 30 minutes.| Product Name | Anti-LC3A/B antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide conjugated with KLH |
| Clonality | polyclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ELISAICC/IFWB |
| ELISA:1:1000-1:10000 WB:1:1000-1:2000 ICC/IF:1:100-1:500 |
|
| Species Reactivity | HumanMouse |
| Concentration | 1mg/ml |
| Purification | Affinity purified |
| Gene Symbol | MAP1LC3A/MAP1LC3B |
|---|---|
| Gene Synonyms | LC3 LC3A ATG8E MAP1ALC3 MAP1BLC3 LC3B ATG8F MAP1LC3B-a MAP1A/1BLC3 |
| Gene Full Name | microtubule associated protein 1 light chain 3 alpha/beta |
| Gene Summary | MAP1A and MAP1B are microtubule-associated proteins which mediate the physical interactions between microtubules and components of the cytoskeleton. MAP1A and MAP1B each consist of a heavy chain subunit and multiple light chain subunits. The protein encoded by this gene is one of the light chain subunits and can associate with either MAP1A or MAP1B. Two transcript variants encoding different isoforms have been found for this gene. The expression of variant 1 is suppressed in many tumor cell lines, suggesting that may be involved in carcinogenesis. [provided by RefSeq, Feb 2012] |
| Alternative Names | ATG8E Autophagy-related protein LC3 A Autophagy-related ubiquitin-like modifier LC3 A LC3 LC3A MAP1 light chain 3 like protein 1 MAP1 light chain 3-like protein 1 MAP1A/1B light chain 3 A MAP1A/MAP1B LC3 A MAP1A/MAP1B light chain 3 A MAP1ALC3 MAP1BLC3 Map1lc3a Microtubule associated proteins 1A/1B light chain 3 Microtubule-associated protein 1 light chain 3 alpha Microtubule-associated proteins 1A and 1B light chain 3 Microtubule-associated proteins 1A/1B light chain 3A MLP3A_HUMAN ATG8F Autophagy-related protein LC3 B Autophagy-related ubiquitin-like modifier LC3 B LC3B LC3II MAP1 light chain 3 like protein 2 MAP1 light chain 3-like protein 2 MAP1A/1BLC3 MAP1A/MAP1B LC3 B MAP1A/MAP1B light chain 3 B MAP1ALC3 MAP1LC3B a Map1lc3b Microtubule associated protein 1 light chain 3 beta Microtubule-associated protein 1 light chain 3 beta Microtubule-associated proteins 1A/1B light chain 3B MLP3B_HUMAN |
| Molecular Weight(MW) | 14kDa（observed：16、20kDa) |
| Cellular Localization | Cytoplasm, Cytoplasmic vesicle, Endomembrane system, Lipid-anchor, autophagosome, autophagosome membrane, cytoskeleton. |
| SwissProt ID | Q9H492&Q9GZQ8 |
|---|
WB
Western blot analysis using LC3A/B antibody against B16(1),A549(2),Nih3t3(3) cell . 12% SDS-PAGE gel.Sample loading: 20μg /lane. Transfer the proteins onto a PVDF membrane (OM790003), and block it with TBST (OM750016) plus skimmed milk powder(OM750084) for one hour. Dilute the primary antibody with the antibody diluent (OM750012) at a ratio of 1:1000, and incubate it overnight at 4°C. Wash the membrane three times with TBST (OM750016), 5 minutes each time. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP (OM643487), at a ratio of 1:20000 and incubate for one hour. Wash the membrane three times with TBST (OM750016) again, 5 minutes each time. Use ECL (OM625701) for luminescence.staining time: 60S.
ELISA
Microtiter plate wells were coated with 100 ng/well coating antigen and incubated with serial dilutions of the antibody. Following HRP-conjugated secondary antibody and substrate development, OD450 was measured. The absorbance decreased progressively with increasing antibody dilution, yielding a sigmoidal titration curve that demonstrates the high antigen-binding activity of the polyclonal antibody.
ICC/IF
Immunofluorescence analysis of NIH3T3 cells using LC3A/B antibody (green). Blue: DAPI fluorescent DNA dye. Red: Actin filaments have been labeled with Omnimabs® 594-Phalloidin.Cells are fixed in 4% paraformaldehyde at room temperature for 15 minutes. Then, they are permeabilized with a PBS (OM750003) solution containing 0.1% Triton X-100(OM750021) at room temperature for 15 minutes. Subsequently, the cells are blocked with 10% non - immune goat serum(OM760028) at room temperature for 1 hour.The cells are incubated overnight at 4°C with the primary antibody diluted 1:100 in PBS. The secondary antibody, Omnimabs® 488 Goat Ant-Rabbit IgG(H&L) (Green,OM643486), is diluted at a ratio of 1:400 and incubated with the cells for 1 hour.Nuclear DNA is labeled with DAPI (Blue,OM643160). F-actin is stained with Omnimabs® 594-Phalloidin (Red,OM750007) diluted 1:100 for 30 minutes.| Application Notes | ELISA:1:1000-1:10000 WB:1:1000-1:2000 ICC/IF:1:100-1:500 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C long term. Avoid freeze / thaw cycle. |
| Storage Buffer | Purified antibody in PBS with 0.05% sodium azide,40% Glycerol. |
