製品の概要

  • 製品名
  • 製品の詳細
    Rabbit polyclonal to LC3B
  • アプリケーション
    適用あり: IHC-P, WB, ICC/IFmore details
  • 種交差性
    交差種: Mouse, Rat, Human
    交差が予測される動物種: Chicken, Cow, Macaque monkey, Zebra finch
  • 免疫原

    Synthetic peptide corresponding to Human LC3B aa 1-100 (N terminal) conjugated to Keyhole Limpet Haemocyanin (KLH). Peptide available as ab87823.
    Database link: Q9GZQ8
    (Peptide available as ab87823, ab87823)

  • ポジティブ・コントロール
    • This antibody gave a positive signal in Human, Mouse and Rat Brain Tissue Lysate. ICC/IF: HeLa cells (+/- chloroquine), HAP1 cells (+/-chloroquine) (HAP1-MAP1LC3B knockout cells used as negative cell line).

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab63817 in the following tested applications.

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

アプリケーション Abreviews 特記事項
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa).
ICC/IF Use a concentration of 1 µg/ml.

ターゲット情報

  • 機能
    Probably involved in formation of autophagosomal vacuoles (autophagosomes).
  • 組織特異性
    Most abundant in heart, brain, skeletal muscle and testis. Little expression observed in liver.
  • 配列類似性
    Belongs to the MAP1 LC3 family.
  • 翻訳後修飾
    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.
  • 細胞内局在
    Cytoplasm > cytoskeleton. Endomembrane system. Cytoplasmic vesicle > autophagosome membrane. LC3-II binds to the autophagic membranes.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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

画像

  • ab63817 staining LC3B in HAP1 cells (wildtype and MAP1LC3B knockout) +/- Chloroquine (50μM, 24 hours). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab63817 at 1μg/ml and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 1/250 dilution (shown in pseudocolor red) followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).



  • Predicted band size : 15 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: LC3B knockout HAP1 cell lysate (20 µg)
    Lane 3: Human brain tissue lysate (20 µg)
    Lane 4: U87MG cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab63817 observed at 16 kDa. Red - loading control, ab8245, observed at 37 kDa.
    ab63817 was shown to recognize LC3B when LC3B knockout samples were used, along with additional cross-reactive bands. Wild-type and LC3B knockout samples were subjected to SDS-PAGE. ab63817 and ab8245 (loading control to GAPDH) were diluted 1 µg/mL and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • ab63817 staining LC3B in HeLa cells +/- Chloroquine (50μM, 24 hours). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab63817 at 1μg/ml and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 1/250 dilution (shown in pseudocolor red) followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • IHC image of LC3B staining in human hippocampus formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab63817, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  • Anti-LC3B antibody (ab63817) at 1 µg/ml + Human brain tissue lysate - total protein (ab29466) at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 15 kDa
    Observed band size : 15 kDa
    Additional bands at : 32 kDa,54 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 2 minutes
  • All lanes : Anti-LC3B antibody (ab63817) at 1 µg/ml

    Lane 1 : Brain (Mouse) Tissue Lysate
    Lane 2 : Brain (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 15 kDa
    Observed band size : 15 kDa
    Additional bands at : 30 kDa,65 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 3 minutes
  • Anti-LC3B antibody (ab63817) at 1 µg/ml + LC3B protein (Human) (ab87944) at 0.1 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 15 kDa


    Exposure time : 2 minutes
  • Immunocytochemical analysis of HeLa cells, labeling LC3B with ab63817 (1/200 with PBS for 16 hours at 22°C). Cells were methanol fixed, immunofluorescently labeled, adn counterstained with DAPI. 

    See Abreview

参考文献

This product has been referenced in:
  • Piao X  et al. Picroside II Shows Protective Functions for Severe Acute Pancreatitis in Rats by Preventing NF-?B-Dependent Autophagy. Oxid Med Cell Longev 2017:7085709 (2017). WB . Read more (PubMed: 28713490) »
  • Lin Q  et al. Cisplatin regulates cell autophagy in endometrial cancer cells via the PI3K/AKT/mTOR signalling pathway. Oncol Lett 13:3567-3571 (2017). Read more (PubMed: 28521459) »

See all 13 Publications for this product

レビューと Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Methanol
Permeabilization
No
Username

Dr. Kirk Mcmanus

Verified customer

投稿 May 10 2013

Thank you for contacting Abcam. I do wish to apologize for the delay in getting this information to you.

We have been in touch with the lab regarding your question of specificity of ab48394 to LC3B. As the immunogen sequence has been conside...

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Thank you for your reply.

I will be pleased to provide an free of charge vial of from a different lot.

Regrettably, this antibody is not in stock at the moment. Another batch is in production and is expected to be available by the...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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