Overview

  • Product name
    Anti-Integrin alpha 2+beta 1 antibody [P1E6]
    See all Integrin alpha 2+beta 1 primary antibodies
  • Description
    Mouse monoclonal [P1E6] to Integrin alpha 2+beta 1
  • Host species
    Mouse
  • Specificity
    ab24697 is directed against the a2 subunit and will precipitate a complex containing a2, including a2ß1 Integrin and neutralizes binding to collagen.
  • Tested applications
    Suitable for: IP, Blocking, Neutralising, Flow Cyt, Inhibition Assay, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Tissue/ cell preparation (Human): HT1080 fibrosarcoma cells.

  • Positive control
    • IF: HepG2 cell line
  • General notes

    This antibody clone is manufactured by Abcam.

    ab24697 is effective in blocking adhesion cells to collagen coated surfaces. It immunoprecipitates the a2 subunit (reducing) or a2 complexed with ß1 from surface labeled cell lysates (non-reducing). In imunofluorescence, this antibody can be used to detect a2 in collagen dependent focal adhesions.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab24697 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 at an assay dependent concentration.
Blocking Use at an assay dependent concentration.
Neutralising Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

(Also PMID - 19780039)

 

 

 

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

Inhibition Assay Use at an assay dependent concentration.
ICC/IF Use a concentration of 5 - 10 µg/ml.

Target

  • Cellular localization
    Cell Membrane; single-pass type I membrane protein
  • Database links
  • Alternative names
    • CD29 antibody
    • CD49B antibody
    • Collagen receptor antibody
    • GPIa antibody
    • GPIIA antibody
    • Integrin alpha 2 antibody
    • Integrin beta 1 antibody
    • ITGA2 antibody
    • ITGB1 antibody
    • Platelet antigen Br antibody
    • Platelet glycoprotein GPIa antibody
    • Platelet glycoprotein Ia antibody
    • Platelet membrane glycoprotein Ia antibody
    • Very late activation protein 2 receptor alpha 2 subunit antibody
    • VLA 2 alpha chain antibody
    • VLA 2 antibody
    • VLA4 antibody
    • VLAA2 antibody
    see all

Images

  • Overlay histogram showing HT1080 cells stained with ab24697 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab24697, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was a goat anti-mouse DyLight® 488 (IgG; H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive result in 80% methanol (5 min) fixed HT1080 cells used under the same conditions.

    Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

  • ICC/IF image of ab24697 stained HepG2 cells. The cells were 4% paraformaldehyde 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 (ab24697, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse 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.

References

This product has been referenced in:
  • Afratis NA  et al. IGF-IR cooperates with ERa to inhibit breast cancer cell aggressiveness by regulating the expression and localisation of ECM molecules. Sci Rep 7:40138 (2017). Read more (PubMed: 28079144) »
  • Chung CH  et al. Aggretin Venom Polypeptide as a Novel Anti-angiogenesis Agent by Targeting Integrin alpha2beta1. Sci Rep 7:43612 (2017). Read more (PubMed: 28252668) »
See all 10 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Question

Please read below the details of customer's complaint and advise.

Antibody code: ab24697 anti integrin alpha2+beta1 antibody [P1E6]
Batch number: GR30919-4
Antibody storage conditions (temperature/reconstitution etc): aliquots , -20
Description of the problem (high background, wrong band size, more bands, no band etc.) no signal at all, no bands. Other antibodies on the same blot gave bands as expected for example anti beta1 (ab24693).
Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.) platelet lysates
Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)
Lysis buf: TBS, NP-40 1%, deoxycholic acid 1%,protease inhibitors cocktail
Heating sample before electrophoresis: 1000C, 5 min
Amount of protein loaded 30μg protein
Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.):
non-reducing, 8% gel, Hepes running buffer
Transfer and blocking conditions (Buffer/time period, Blocking agent etc.) no problem in transferring because as I mention before the blot worked well with other antibodies.
Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
Anti integrin alpha2 (P1E6) ab24697. I tried 2 dilutions: 1:1000 and 1:500. 2 hours incubation. 3 times washes in TBS-T.
Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step): peroxidase-conjugated goat anti-mouse IgG (Jackson ImmunoResearch)
Detection method (ECL, ECLPlus etc.): SuperSignal West Pico Chelimuniescent Substrate
Positive and negative controls used (please specify) - total normal platelet lysates
Optimization attempts (problem solving)
How many times have you tried the Western? Twice with 2 different blots
Have you run a "No Primary" control? No
Do you obtain the same results every time? e.g. are the background bands always in the same place? irrelevant
What steps have you altered? I tried to put twice the amount of the primary antibody. I checked if the blot still reacts with other antibodies for example anti calnexin and got the expected results.
Thanks in advance for your assistance and reply.

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Answer

Thank you for contacting us.
I am sorry to hear that this antibody is not providing satisfactory results in Western Blot.
However, as indicated on the datasheet (www.abcam.com/ab24697), this antibody has not been tested in Western Blot and we cannot guarantee it to work in this application.
I would suggest to try an incubation of the antibody overnight at 4ºC.
Maybe the antibody is just not suitable for Western Blot or not suitable for the detection of denaturated protein. If so, I would suggest to run a Western Blot in non-denaturing conditions.
I hope this is helpful.

Read More

Answer

Thank you for your enquiry. Ab24697 is directed against integrin alpha2 and neutralizes binding to collagen and laminin. The following references will be useful for you: Wayne, E.A., and Carter, W.G., 1987, J Cell Biol, 105:1873-1884. Wayne, et al., 1990, J Cell Biol, 110:1387-1404. I hope this information helps. Please do not hesitate to contact us if you need anything further.

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