Overview

  • Product nameAnti-LC3A/B antibodySee all LC3A/B primary antibodies ...
  • Description
    Rabbit polyclonal to LC3A/B
  • Tested applicationsWB, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Cow, Zebrafish
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human LC3A/B.

    (Peptide available as ab99367.)

  • Positive control
    • This antibody gave a positive signal in SHSY-5Y and SK N BE Whole Cell Lysates, and in mouse and rat brain tissue lysates. IHC-P: FFPE human hippocampus tissue sections.

Properties

Applications

Our Abpromise guarantee covers the use of ab84936 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 Use a concentration of 1 µg/ml. Detects a band of approximately 18 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human LC3A/B peptide (ab99367).
ICC/IF Use a concentration of 10 µg/ml.
IHC-P Use a concentration of 0.5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

Anti-LC3A/B antibody images

  • IHC image of LC3A/B staining in human hippocampus formalin fixed paraffin embedded tissue section, performed on a Leica Bond 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 ab84936, 0.5µg/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.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • All lanes : Anti-LC3A/B antibody (ab84936) at 1 µg/ml

    Lane 1 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
    Lane 2 : SK N BE (Human neuroblastoma) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    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 : 18 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 45-70 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 12 minutes
  • ICC/IF image of ab84936 stained PC12 cells. The cells were 4% formaldehyde (10 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 (ab84936, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.

  • All lanes : Anti-LC3A/B antibody (ab84936) 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 : 18 kDa,30 kDa,50 kDa,70 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 4 minutes

References for Anti-LC3A/B antibody (ab84936)

ab84936 has not yet been referenced specifically in any publications.

Product Wall

Abreviews
Application Western blot
Sample Human Cell lysate - whole cell (HCT116 colon cancer cell line)
Loading amount 30 µg
Specification HCT116 colon cancer cell line
Gel Running Conditions Reduced Denaturing (13,5%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Christian Marx

Verified customer

Submitted Feb 13 2013

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