Overview

  • Product nameAnti-Tbx3 antibodySee all Tbx3 primary antibodies ...
  • Description
    Rabbit polyclonal to Tbx3
  • Tested applicationsICC/IF, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Chicken
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Tbx3.

  • Positive control
    • This antibody gave a positive signal in nuclear extracts from the following cell lines: HepG2; MCF7. IHC-P (FFPE): Human Placenta (Normal) tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab99302 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
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 79 kDa (predicted molecular weight: 79 kDa).
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

Anti-Tbx3 antibody images

  • IHC image of Tbx3 staining in Human Placenta 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 ab99302, 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.

  • All lanes : Anti-Tbx3 antibody (ab99302) at 1 µg/ml

    Lane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Nuclear Lysate
    Lane 2 : MCF7 (Human breast adenocarcinoma cell line) Nuclear Lysate (ab14860)

    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 : 79 kDa
    Observed band size : 79 kDa
    Additional bands at : 22 kDa,77 kDa (possible isoform). We are unsure as to the identity of these extra bands.

    Exposure time : 3 minutes
    Ab99302 should recognise all three isoforms of Tbx3. We are probably observing Isoform I and Isoform II on this Western Blot image. The molecular weights of Isoform I and Isoform II are 77- and 79-kDa respectively.
  • ICC/IF image of ab99302 stained HepG2 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 ab99302 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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. This antibody also gave a positive result in formaldehyde fixed (4%, 10min) HepG2 cells at 5ug/ml.

References for Anti-Tbx3 antibody (ab99302)

ab99302 has not yet been referenced specifically in any publications.

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - other (E11.5 embryonic limb bud)
Loading amount 10 µg
Specification E11.5 embryonic limb bud
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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Submitted Nov 03 2011

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