Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-PAI1 antibody [EPR17795] - BSA and Azide free (ab250924)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR17795] to PAI1 - BSA and Azide free
  • Suitable for: ICC, IP, Flow Cyt, WB
  • Reacts with: Human

Overview

  • Product name

    Anti-PAI1 antibody [EPR17795] - BSA and Azide free
    See all PAI1 primary antibodies

  • Description

    Rabbit monoclonal [EPR17795] to PAI1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC, IP, Flow Cyt, WBmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: A549, HepG2 and HUVEC cell lysates; Human fetal liver and spleen lysates; Human PAI1 full length protein. Flow Cyt: HepG2 cells. ICC: HepG2 cells. IP: HepG2 cells.

  • General notes

    Ab250924 is the carrier-free version of ab187262. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab250924 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Western blot - Anti-PAI1 antibody [EPR17795] - BSA and Azide free (ab250924)
    Western blot - Anti-PAI1 antibody [EPR17795] - BSA and Azide free (ab250924)
    All lanes : Anti-PAI1 antibody [EPR17795] (ab187262) at 1/1000 dilution

    Lane 1 : Wild-type A549 cell lysate
    Lane 2 : SERPINE1 knockout A549 cell lysate
    Lane 3 : HUVEC cell lysate
    Lane 4 : HEK-293 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 45 kDa
    Observed band size: 48 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab187262).

    Lanes 1 - 4: Merged signal (red and green). Green - ab187262 observed at 48 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

    ab187262 was shown to react with PAI1 in wild-type A549 cells in Western blot. The band observed in the edited lysate lane above 45 kDa is likely to represent SERPINE1 with an insertion. This has not been investigated further. Wild-type A549 and SERPINE1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab187262 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

  • Immunocytochemistry - Anti-PAI1 antibody [EPR17795] - BSA and Azide free (ab250924)
    Immunocytochemistry - Anti-PAI1 antibody [EPR17795] - BSA and Azide free (ab250924)

    This data was developed using ab187262, the same antibody clone in a different buffer formulation.

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma) cells labeling PAI1 with ab187262 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Cytoplasm staining on HepG2 cells was observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).The negative controls are as follows:
    -ve control 1: ab187262 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.
  • Flow Cytometry - Anti-PAI1 antibody [EPR17795] - BSA and Azide free (ab250924)
    Flow Cytometry - Anti-PAI1 antibody [EPR17795] - BSA and Azide free (ab250924)
    This data was developed using ab187262, the same antibody clone in a different buffer formulation.ab187262 staining PAI1 in the human cell line HepG2 (human hepatocellular carcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/60. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody. Isoytype control: Rabbit monoclonal IgG (Black) Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)
  • Immunoprecipitation - Anti-PAI1 antibody [EPR17795] - BSA and Azide free (ab250924)
    Immunoprecipitation - Anti-PAI1 antibody [EPR17795] - BSA and Azide free (ab250924)
    This data was developed using ab187262, the same antibody clone in a different buffer formulation.PAI1 was immunoprecipitated from 1mg of HepG2 (Human liver hepatocellular carcinoma) whole cell lysate with ab187262 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab187262 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1500 dilution. Lane 1: HepG2 whole cell lysate 10 µg (Input). Lane 2: ab187262 IP in HepG2 whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab187262 in HepG2 whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST. Exposure time: 30 seconds.
  • Western blot - Anti-PAI1 antibody [EPR17795] - BSA and Azide free (ab250924)
    Western blot - Anti-PAI1 antibody [EPR17795] - BSA and Azide free (ab250924)
    All lanes : Anti-PAI1 antibody [EPR17795] (ab187262) at 1/1000 dilution

    Lane 1 : Human fetal liver lysate
    Lane 2 : Human fetal spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 45 kDa
    Observed band size: 45 kDa


    Exposure time: 3 minutes


    This data was developed using ab187262, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry - Anti-PAI1 antibody [EPR17795] - BSA and Azide free (ab250924)
    Immunocytochemistry - Anti-PAI1 antibody [EPR17795] - BSA and Azide free (ab250924)
    This data was developed using ab187262, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HT1080 (Human fibrosarcoma cells) cells labeling PAI1 with ab187262 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Cytoplasm staining on HT1080 cells was observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).The negative controls are as follows:
    -ve control 1: ab187262 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.
  • Western blot - Anti-PAI1 antibody [EPR17795] - BSA and Azide free (ab250924)
    Western blot - Anti-PAI1 antibody [EPR17795] - BSA and Azide free (ab250924)
    Anti-PAI1 antibody [EPR17795] (ab187262) at 1/5000 dilution + HepG2 (Human liver hepatocellular carcinoma) whole cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 45 kDa
    Observed band size: 45 kDa


    Exposure time: 1 minute


    This data was developed using ab187262, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-PAI1 antibody [EPR17795] - BSA and Azide free (ab250924)
    Western blot - Anti-PAI1 antibody [EPR17795] - BSA and Azide free (ab250924)
    Anti-PAI1 antibody [EPR17795] (ab187262) at 1/5000 dilution + Human PAI1 full length protein at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 45 kDa
    Observed band size: 70 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    This data was developed using ab187262, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Anti-PAI1 antibody [EPR17795] - BSA and Azide free (ab250924)
    Anti-PAI1 antibody [EPR17795] - BSA and Azide free (ab250924)

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