Overview

  • Product name
    Anti-LC3B antibody - N-terminal
    See all LC3B primary antibodies
  • Description
    Rabbit polyclonal to LC3B - N-terminal
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, Flow Cyt, ICC/IF, IP, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Pig
    Predicted to work with: Cow, Chimpanzee, Zebrafish, Rhesus monkey, Medaka fish
  • Immunogen

    Synthetic peptide within Human LC3B (N terminal). The exact sequence is proprietary. (Carrier-protein conjugated).
    Database link: Q9GZQ8

  • Positive control
    • ICC/IF: HeLa cells untreated and treated with 50µM Chloroquine for 24 hr; Hep G2 cells untreated and treated with 3 µM Thapsigargin for 12 hrs. WB: Huh7 whole cell lysate; NTERA-2 cl.D1 [NT2/D1], PC-3, U-87 MG and SK-N-SH whole cell extracts; Mouse and rat brain lysates; Hep G2 cell lysate untreated and treated with 3 µM Thapsigargin; HeLa whole cell extract untreated and treated with 50µM Chloroquine for 24 hr. IHC-P: Mouse brain tissue. IP: U-87 MG whole cell extract. Flow Cyt: HeLa cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab229327 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/500 - 1/3000. Predicted molecular weight: 15 kDa.
Flow Cyt 1/50 - 1/200.
ICC/IF 1/100 - 1/1000.
IP 1/100 - 1/500.
IHC-P 1/100 - 1/1000.

Target

  • Function
    Probably involved in formation of autophagosomal vacuoles (autophagosomes).
  • Tissue specificity
    Most abundant in heart, brain, skeletal muscle and testis. Little expression observed in liver.
  • Sequence similarities
    Belongs to the MAP1 LC3 family.
  • Post-translational
    modifications
    The precursor molecule is cleaved by APG4B/ATG4B to form LC3-I. This is activated by APG7L/ATG7, transferred to ATG3 and conjugated to phospholipid to form LC3-II.
  • Cellular localization
    Cytoplasm > cytoskeleton. Endomembrane system. Cytoplasmic vesicle > autophagosome membrane. LC3-II binds to the autophagic membranes.
  • Information by UniProt
  • Database links
  • Alternative names
    • ATG8F antibody
    • Autophagy-related protein LC3 B antibody
    • Autophagy-related ubiquitin-like modifier LC3 B antibody
    • LC3B antibody
    • LC3II antibody
    • MAP1 light chain 3 like protein 2 antibody
    • MAP1 light chain 3-like protein 2 antibody
    • MAP1A/1BLC3 antibody
    • MAP1A/MAP1B LC3 B antibody
    • MAP1A/MAP1B light chain 3 B antibody
    • MAP1ALC3 antibody
    • MAP1LC3B a antibody
    • Map1lc3b antibody
    • Microtubule associated protein 1 light chain 3 beta antibody
    • Microtubule-associated protein 1 light chain 3 beta antibody
    • Microtubule-associated proteins 1A/1B light chain 3B antibody
    • MLP3B_HUMAN antibody
    see all

Images

  • All lanes : Anti-LC3B antibody - N-terminal (ab229327) at 1/1500 dilution

    Lane 1 : Huh7 whole cell lysate (untreated)
    Lane 2 : Huh7 whole cell lysate (3 µM Thapsigargin treatment for 12 hr)

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-rabbit IgG

    Predicted band size: 15 kDa

  • 4% paraformaldehyde-fixed HeLa (human epithelial cell line from cervix adenocarcinoma) cells, mock treated (left panel) or treated with 50 μM Chloroquine for 24 hr (right panel), stained for LC3B (green) using ab229327 at 1/2000 dilution in ICC/IF.

    Red: alpha Tubulin, a cytoskeleton marker, stained by alpha Tubulin antibody at 1/1000 dilution.

    Blue: Hoechst 33342 staining.

  • Paraffin-embedded mouse brain tissue stained for LC3B using ab229327 at 1/500 dilution in immunohistochemical analysis.

  • All lanes : Anti-LC3B antibody - N-terminal (ab229327) at 1/1500 dilution

    Lane 1 : Huh7 whole cell lysate (un-infected)
    Lane 2 : Huh7 whole cell lysate (HCV-infected)

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-rabbit IgG

    Predicted band size: 15 kDa

  • All lanes : Anti-LC3B antibody - N-terminal (ab229327) at 1/1000 dilution

    Lane 1 : NTERA-2 cl.D1 [NT2/D1] (human malignant pluripotent embryonic carcinoma cell line) whole cell extract
    Lane 2 : PC-3 (human prostate adenocarcinoma cell line) whole cell extract
    Lane 3 : U-87 MG (human glioblastoma-astrocytoma epithelial cell line) whole cell extract
    Lane 4 : SK-N-SH (human neuroblastoma cell line) whole cell extract

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-rabbit IgG

    Predicted band size: 15 kDa



    15% SDS-PAGE gel.

  • Ice-cold methanol-fixed, ice-cold acetone permeabilized HepG2 (human liver hepatocellular carcinoma cell line) cells, mock treated (left panels) or treated with with 3 μM Thapsigargin for 12 hrs (right panels), stained for LC3B (green) using ab229327 at 1/500 dilution in ICC/IF.

    Blue: Hoechst 33342 staining.

  • LC3B was immunoprecipitated from U-87 MG (human glioblastoma-astrocytoma epithelial cell line) whole cell extract (1 mg for IP, 20% of IP loaded) with 5 µg ab229327. Western blot was performed from the immunoprecipitate using ab229327. Anti-Rabbit IgG was used as a secondary reagent.

    Lane 1: U-87 MG whole cell extract.

    Lane 2: Control IgG IP in  U-87 MG  whole cell extract.

    Lane 3: ab229327 IP in  U-87 MG  whole cell extract.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling LC3B with ab229327 at 1/100 dilution (blue) compared with an unlabelled sample (brown).

    Acquisition of >20,000 events were collected using Argon ion laser (488nm) and 525/30 bandpass filter.

  • 4% paraformaldehyde-fixed HeLa (human epithelial cell line from cervix adenocarcinoma) cells, mock treated (left panels) or treated with 50 μM Chloroquine for 24 hr (right panels), stained for LC3B (green) using ab229327 at 1/200 dilution in ICC/IF.

    Red: phalloidin, a cytoskeleton marker, at 1/50 dilution.

    Blue: Hoechst 33342 staining.

  • Anti-LC3B antibody - N-terminal (ab229327) at 1/1000 dilution + Mouse brain lysate at 50 µg

    Secondary
    HRP-conjugated anti-rabbit IgG

    Predicted band size: 15 kDa



    15% SDS-PAGE gel.

  • Anti-LC3B antibody - N-terminal (ab229327) at 1/1000 dilution + Rat brain lysate at 50 µg

    Secondary
    HRP-conjugated anti-rabbit IgG

    Predicted band size: 15 kDa



    15% SDS-PAGE gel.

  • All lanes : Anti-LC3B antibody - N-terminal (ab229327) at 1/1000 dilution

    Lane 1 : Untreated HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate
    Lane 2 : HepG2 whole cell lysate treated with 3 µM thapsigargin for 12 hours

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-rabbit IgG

    Predicted band size: 15 kDa



    Lower panel: Beta-actin antibody at 1/20000 dilution.

    15% SDS-PAGE gel.

  • All lanes : Anti-LC3B antibody - N-terminal (ab229327) at 1/1000 dilution

    Lane 1 : Untreated HepG2 (human liver hepatocellular carcinoma cell line) whole cell extract
    Lane 2 : HepG2 whole cell extract treated with 3 µM thapsigargin for 16 hours
    Lane 3 : HepG2 whole cell extract treated with 3 µM thapsigargin for 24 hours

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-rabbit IgG

    Predicted band size: 15 kDa



    15% SDS-PAGE gel.

  • All lanes : Anti-LC3B antibody - N-terminal (ab229327) at 1/2500 dilution

    Lane 1 : Untreated HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell extract
    Lane 2 : HeLa whole cell extract treated with 50µM Chloroquine for 24 hours

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-rabbit IgG

    Predicted band size: 15 kDa



    15% SDS-PAGE gel.

References

ab229327 has not yet been referenced specifically in any publications.

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