MAP1LC3A Recombinant Rabbit Monoclonal Antibody [ST47-03]
Catalog# ET1609-26
MAP1LC3A Recombinant Rabbit Monoclonal Antibody [ST47-03]
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WB
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IF-Cell
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FC
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IP
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
MAP1LC3A Recombinant Rabbit Monoclonal Antibody [ST47-03]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human MAP1LC3A aa 81-121 / 121.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, FC, IP
Molecular Weight
Predicted band size: 14 kDa
Positive Control
HeLa treated with 50μM Chloroquine for 18 hours cell lysate, C2C12 cell lysate, C2C12 treated with 50μM Chloroquine for 18 hours cell lysate, C6 cell lysate, C6 treated with 50μM Chloroquine for 18 hours cell lysate, SH-SY5Y, Neuro-2a, C6.
Conjugation
unconjugated
Clone Number
ST47-03
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:1,000-1:2,000
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IF-Cell
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1:100
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FC
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1:1,000
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IP
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1-2μg/sample
Target
Function
Microtubules, the primary component of the cytoskeletal network, interact with proteins called microtubule-associated proteins (MAPs). The microtubule-associated proteins can be divided into two groups, structural and dynamic. The structural microtubule-associated proteins, MAP-1A, MAP-1B, MAP-2A, MAP-2B and MAP-2C, stimulate tubulin assembly, enhance micro-tubule stability and influence the spatial distribution of microtubules within cells. Both MAP-1 and, to a greater extent, MAP-2 have been implicated as agents of microtubule depolymerization by suppressing the dynamic instability of the microtubules. The suppression of microtubule dynamic instability by the MAP proteins is thought to be associated with phosphorylation of the MAPs.
Background References
1. Kim JY et al. Expression of autophagy-related proteins according to androgen receptor and HER-2 status in estrogen receptor-negative breast cancer. PLoS One 9:e105666 (2014).
2. Jung CH et al. ULK-Atg13-FIP200 complexes mediate mTOR signaling to the autophagy machinery. Mol Biol Cell 20:1992-2003 (2009).
Sequence Similarity
Belongs to the ATG8 family.
Tissue Specificity
Most abundant in heart, brain, liver, skeletal muscle and testis but absent in thymus and peripheral blood leukocytes.
Post-translational Modification
The precursor molecule is cleaved by ATG4B to form the cytosolic form, LC3-I. This is activated by APG7L/ATG7, transferred to ATG3 and conjugated to phospholipid to form the membrane-bound form, LC3-II.; The Legionella effector RavZ is a deconjugating enzyme that produces an ATG8 product that would be resistant to reconjugation by the host machinery due to the cleavage of the reactive C-terminal glycine.; Phosphorylation at Ser-12 by PKA inhibits conjugation to phosphatidylethanolamine (PE). Interaction with MAPK15 reduces the inhibitory phosphorylation and increases autophagy activity.
Subcellular Location
Cytoplasm, Endomembrane system, Cytoplasmic vesicle, Microtubule, Membrane.
Synonyms
ATG8E antibody
Autophagy-related protein LC3 A antibody
Autophagy-related ubiquitin-like modifier LC3 A antibody
LC3 antibody
LC3A antibody
MAP1 light chain 3 like protein 1 antibody
MAP1 light chain 3-like protein 1 antibody
MAP1A/1B light chain 3 A antibody
MAP1A/MAP1B LC3 A antibody
MAP1A/MAP1B light chain 3 A antibody
ExpandImages
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☑ Cell treatment (CT)
Western blot analysis of MAP1LC3A on different lysates with Rabbit anti-MAP1LC3A antibody (ET1609-26) at 1/1,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: HeLa treated with 50μM Chloroquine for 18 hours cell lysate
Lane 3: C2C12 cell lysate
Lane 4: C2C12 treated with 50μM Chloroquine for 18 hours cell lysate
Lane 5: C6 cell lysate
Lane 6: C6 treated with 50μM Chloroquine for 18 hours cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 14 kDa
Observed band size: 14/16 kDa
Exposure time: 3 minutes; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1609-26) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of SH-SY5Y cells labeling MAP1LC3A with Rabbit anti-MAP1LC3A antibody (ET1609-26) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-MAP1LC3A antibody (ET1609-26) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunocytochemistry analysis of Neuro-2a cells labeling MAP1LC3A with Rabbit anti-MAP1LC3A antibody (ET1609-26) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-MAP1LC3A antibody (ET1609-26) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunocytochemistry analysis of C6 cells labeling MAP1LC3A with Rabbit anti-MAP1LC3A antibody (ET1609-26) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-MAP1LC3A antibody (ET1609-26) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Flow cytometric analysis of SH-SY5Y cells labeling MAP1LC3A.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET1609-26, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
Flow cytometric analysis of Neuro-2a cells labeling MAP1LC3A.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET1609-26, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
Flow cytometric analysis of C6 cells labeling MAP1LC3A.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET1609-26, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
MAP1LC3A was immunoprecipitated from 0.2 mg C6 cell lysate with ET1609-26 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1609-26 at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: C6 cell lysate (input)
Lane 2: ET1609-26 IP in C6 cell lysate
Lane 3: Rabbit IgG instead of ET1609-26 in C6 cell lysate
Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 3 minutes; ECL: K1801
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Citation
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Dynamin-Related Protein 1 Is Involved in Mitochondrial Damage, Defective Mitophagy, and NLRP3 Inflammasome Activation Induced by MSU Crystals
Journal: Oxidative Medicine And Cellular Longevity
DOI:
IF: 7.310
Application: WB
Reactivity: Mouse
Publish date: 2022 Oct
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