概述

  • 产品名称Anti-LC3B抗体
    参阅全部 LC3B 一抗
  • 描述
    兔多克隆抗体to LC3B
  • 经测试应用适用于: IHC-P, WB, ICC/IFmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
    预测可用于: 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)

  • 阳性对照
    • This antibody gave a positive signal in Human, Mouse and Rat Brain Tissue Lysate.

性能

应用

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.

应用 Ab评论 说明
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 5 µg/ml.

靶标

  • 功能Probably involved in formation of autophagosomal vacuoles (autophagosomes).
  • 组织特异性Most abundant in heart, brain, skeletal muscle and testis. Little expression observed in liver.
  • 序列相似性Belongs to the MAP1 LC3 family.
  • 翻译后修饰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.
  • 细胞定位Cytoplasm > cytoskeleton. Endomembrane system. Cytoplasmic vesicle > autophagosome membrane. LC3-II binds to the autophagic membranes.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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

Anti-LC3B antibody 图像



  • 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) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • ICC/IF image of ab63817 stained PC12 cells. The cells were 100% Methanol fixed (5 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab63817, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
  • 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

Anti-LC3B antibody (ab63817)参考文献

This product has been referenced in:
  • Wang J  et al. Aberrant expression of Beclin-1 and LC3 correlates with poor prognosis of human hypopharyngeal squamous cell carcinoma. PLoS One 8:e69038 (2013). WB, IHC . Read more (PubMed: 23935917) »
  • Huang J  et al. Effect of a low-protein diet supplemented with ketoacids on skeletal muscle atrophy and autophagy in rats with type 2 diabetic nephropathy. PLoS One 8:e81464 (2013). WB ; Rat . Read more (PubMed: 24303049) »

See all 3 Publications for this product

Product Wall

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

提交于 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...

Read More

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...

Read More

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