Overview

  • Product name

  • Description

    Rabbit polyclonal to Zyxin
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Rat, Human
    Predicted to work with: Mouse, Dog, Rhesus monkey
  • Immunogen

    Synthetic peptide corresponding to Human Zyxin aa 450-550 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab71841)

  • Positive control

Properties

Applications

Our Abpromise guarantee covers the use of ab71842 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
IP Use a concentration of 5 µg/ml.
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 75 kDa (predicted molecular weight: 61 kDa).

Target

  • Relevance

    Zyxin binds alpha-actinin and the CRP proteins and is implicated in the regulation and modulation of actin filaments organization, cell adhesion, cell motility, mitosis, and signal transduction. Zyxin may shuttle between focal adhesion plaques and the nucleus. It has been suggested that zyxin is involved in gene transcription by its association with transcription factors in the nucleus.
  • Cellular localization

    Cytoplasmic, cytoskeletal and Nuclear.
  • Database links

  • Alternative names

    • ESP 2 antibody
    • ESP2 antibody
    • HED 2 antibody
    • HED2 antibody
    • ZYX antibody
    • ZYX protein antibody
    • Zyxin 2 antibody
    • Zyxin2 antibody
    see all

Images

  • All lanes : Anti-Zyxin antibody (ab71842) at 1 µg/ml

    Lane 1 : SK N SH (Human neuroblastoma) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 3 : A498 (Human Kidney Carcinoma) Whole Cell Lysate
    Lane 4 : U-87 MG (Human glioblastoma astrocytoma) Whole Cell Lysate
    Lane 5 : DU 145 (Human prostate carcinoma cell line) Whole Cell Lysate
    Lane 6 : Testis (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size: 61 kDa
    Observed band size: 75 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 100 kDa, 48 kDa. We are unsure as to the identity of these extra bands.

  • Zyxin was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Zyxin and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab71842.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 75kDa; Zyxin
  • ICC/IF image of ab71842 stained HepG2 cells. The cells were 4% PFA fixed (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 (ab71842, 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. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa cells at 5µg/ml, and in 100% Methanol fixed (5 min) Hela, Hek293, HepG2, and MCF-7 cells at 5µg/ml.

References

This product has been referenced in:

See all 7 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293T cells)
Loading amount
25 µg
Specification
HEK293T cells
Gel Running Conditions
Reduced Denaturing (7% SDS-PAGE gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Jan 24 2013

Answer

DISCOUNT CODE: xxx for ab19347
DISCOUNT CODE: xxx for ab71842
Expiration date: xxx

I am very pleased to hear you would like to accept our offer and test ab19347 and ab71842in chicken and/or IHC-Fr. Each code will give you: 1 free PRIMARY ANTIBODYbefore the expiration date. To redeem this offer, please submit an Abreview for chicken and/or IHC-Frand include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Formaldehyde
Permeabilization
Yes - 0.1% Triton x-100 in PBS
Blocking step
Milk as blocking agent for 40 minute(s) · Concentration: 10% · Temperature: 23°C

Mrs. Merly Saare

Verified customer

Submitted Feb 18 2010

Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa, Jurkat)
Loading amount
100 µg
Specification
HeLa, Jurkat
Gel Running Conditions
Reduced Denaturing (9%)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 18°C

Mrs. Merly Saare

Verified customer

Submitted Jan 15 2010

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