LC3B-Specific Polyclonal antibody

LC3B-Specific Polyclonal Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat and More (3)

Applications

WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Conjugate

Unconjugated

Publications

311

Reviews (2)

Cat no : 18725-1-AP

Synonyms

LC3, MAP1LC3B, MAP1A/1BLC3, MAP1 light chain 3-like protein 2, LC3B

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Validation Data Gallery

• 

  WB analysis of human brain using 18725-1-AP

  human brain tissue were subjected to SDS PAGE followed by western blot with 18725-1-AP (LC3B-Specific antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of TN treated Hela using 18725-1-AP

  TN treated Hela cells were subjected to SDS PAGE followed by western blot with 18725-1-AP (LC3B-Specific antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of A549 using 18725-1-AP

  A549 cells were subjected to SDS PAGE followed by western blot with 18725-1-AP (LC3B-Specific antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 18725-1-AP

  HepG2 cells were subjected to SDS PAGE followed by western blot with 18725-1-AP (LC3B-Specific antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of mouse brain using 18725-1-AP

  mouse brain tissue were subjected to SDS PAGE followed by western blot with 18725-1-AP (LC3B-Specific antibody) at dilution of 1:300 incubated at room temperature for 1.5 hours.

• 

  WB analysis of MCF-7 using 18725-1-AP

  MCF7 cells were subjected to SDS PAGE followed by western blot with 18725-1-AP (LC3B-Specific antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

• 

  WB analysis of UV treated HEK-293 using 18725-1-AP

  UV treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 18725-1-AP (LC3B-Specific antibody) at dilution of 1:300 incubated at room temperature for 1.5 hours.

• 

  IHC staining of mouse brain using 18725-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 18725-1-AP (LC3B-Specific antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse brain using 18725-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 18725-1-AP (LC3B-Specific antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse brain using 18725-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 18725-1-AP (LC3B-Specific antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat brain using 18725-1-AP

  Immunohistochemical analysis of paraffin-embedded rat brain tissue slide using 18725-1-AP (LC3B-Specific antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat brain using 18725-1-AP

  Immunohistochemical analysis of paraffin-embedded rat brain tissue slide using 18725-1-AP (LC3B-Specific antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse brain using 18725-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 18725-1-AP (LC3B-Specific antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat brain using 18725-1-AP

  Immunohistochemical analysis of paraffin-embedded rat brain tissue slide using 18725-1-AP (LC3B-Specific antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat brain using 18725-1-AP

  Immunohistochemical analysis of paraffin-embedded rat brain tissue slide using 18725-1-AP (LC3B-Specific antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of HepG2 using 18725-1-AP

  Immunofluorescent analysis of -20℃ Ethanol fixed CQ treated HepG2 cells (left) and untreated HepG2 cells (right) using 18725-1-AP (LC3B-Specific antibody) at dilution of 1:50 and Alexa Fluor 488-conjugated Goat Anti-Rabbit IgG(H+L). The cytoskeleton was labelled in red with 66031-1-Ig (alpha tubulin).

• 

  IF Staining of HeLa using 18725-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed Chloroquine treated HeLa cells using LC3B-Specific antibody (18725-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L), CL594-Phalloidin (red).

• 

  IF Staining of HepG2 using 18725-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed CQ treated HepG2 cells using 18725-1-AP (LC3B-Specific antibody) at dilution of 1:200 and CoraLite488-Conjugated Goat Anti-Rabbit IgG(H+L).

• 

  IF Staining of HepG2 using 18725-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed HepG2 cells using 18725-1-AP (LC3B-Specific antibody) at dilution of 1:200 and CoraLite488-Conjugated Goat Anti-Rabbit IgG(H+L).

• 

  FC experiment of HeLa using 18725-1-AP

  1X10^6 HeLa cells were intracellularly stained with 0.4 ug Anti-Human LC3B-Specific (18725-1-AP) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Control Antibody. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Tested Applications

• Positive WB detected in: human brain tissue, MCF-7 cells, TN treated Hela, A549 cells, UV treated HEK-293, mouse brain tissue, HepG2 cells
• Positive IHC detected in: mouse brain tissue, rat brain tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: Chloroquine treated HeLa cells, Chloroquine treated HepG2 cells
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:300-1:1000
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 1 publications below
• WB: See 253 publications below
• IHC: See 44 publications below
• IF: See 88 publications below
• IP: See 3 publications below

Product Information

18725-1-AP targets LC3B-Specific in WB, IHC, IF/ICC, FC (Intra), IP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat, pig, zebrafish, bovine
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: Peptide
• Full Name: microtubule-associated protein 1 light chain 3 beta
• Calculated Molecular Weight: 15 kDa
• Observed Molecular Weight: 15 kDa, 18 kDa
• GenBank Accession Number: NM_022818
• Gene Symbol: LC3B
• Gene ID (NCBI): 81631
• RRID: AB_2137745
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

LC3B, also named as MAP1LC3B, MAP1A/1BLC3, belongs to the MAP1 LC3 family. It is a subunit of neuronal microtubule-associated MAP1A and MAP1B proteins, which are involved in microtubule assembly and important for neurogenesis. In cell biology, autophagy, or autophagocytosis, is a catabolic process involving the degradation of a cell's own components through the lysosomalmachinery. It is a major mechanism by which a starving cell reallocates nutrients from unnecessary processes to more-essential processes. Two forms of LC3, called LC3-I (17-19kd) and -II(14-16kd), were produced post-translationally in various cells. LC3-I is cytosolic, whereas LC3-II is membrane bound. The precursor molecule is cleaved by APG4B/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 amount of LC3-II is correlated with the extent of autophagosome formation. LC3-II is the first mammalian protein identified that specifically associates with autophagosome membranes. MAP1LC3 has 3 isoforms MAP1LC3A, MAP1LC3B and MAP1LC3C. MAP1LC3A and MAP1LC3C are produced by the proteolytic cleavage after the conserved C-terminal Gly residue, like their rat counterpart, MAP1LC3B does not undergo C-terminal cleavage and exists in a single modified form. This antibody is specific to LC3B.

Publications

Species	Application	Title
rat	IHC	Am J Respir Crit Care Med; Acid Sphingomyelinase Inhibition Attenuates Cell Death in Mechanically-Ventilated Newborn Rat Lung.; Authors - Behzad Yeganeh
mouse	WB	Nat Microbiol; Chloroquine ameliorates carbon tetrachloride-induced acute liver injury in mice via the concomitant inhibition of inflammation and induction of apoptosis; Authors - Tomas Doyle
mouse	WB	ACS Nano; Nanoenabled Disruption of Multiple Barriers in Antigen Cross-Presentation of Dendritic Cells via Calcium Interference for Enhanced Chemo-Immunotherapy.; Authors - Jingyi An
human	WB	Autophagy; TRIM59 regulates autophagy through modulating both the transcription and the ubiquitination of BECN1.; Authors - Tianyu Han
human	WB	Autophagy; STYK1 promotes autophagy through enhancing the assembly of autophagy-specific class III phosphatidylinositol 3-kinase complex I.; Authors - Cefan Zhou
human	WB	Mater Today Bio; Carbon dots enhance extracellular matrix secretion for dentin-pulp complex regeneration through PI3K/Akt/mTOR pathway-mediated activation of autophagy.; Authors - Lili Liu

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Azita (Verified Customer) (06-02-2021)

Western blot analysis using LC3B-Specific Polyclonal antibody in NSC34 cell line at dilution of 1:300. Immunohistochemistry labelling of primary rat cortical neurons using LC3B-Specific Polyclonal antibody at dilution of 1:500 showed strong labelling.

• Applications: Western Blot, Immunohistochemistry
• Primary Antibody Dilution: 1:300 (WB), 1:500 (IHC)
• Cell Tissue Type: NSC34

FH

Tanusree (Verified Customer) (12-03-2019)

The antibody works great in Western blotting analysis.

• Applications: Western Blot,
• Primary Antibody Dilution: 1:500
• Cell Tissue Type: Mouse brain

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