Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-COL18A1 antibody [EPR23481-240] - BSA and Azide free (ab275394)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23481-240] to COL18A1 - BSA and Azide free
  • Suitable for: IP, ICC, Flow Cyt, WB
  • Reacts with: Human

Overview

  • Product name

    Anti-COL18A1 antibody [EPR23481-240] - BSA and Azide free
    See all COL18A1 primary antibodies

  • Description

    Rabbit monoclonal [EPR23481-240] to COL18A1 - BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    This antibody does not recognise Endostatin.

    This antibody ab275394 [EPR23481-240] is a distinct clone from ab275750 [EPR23481-21].

  • Tested applications

    Suitable for: IP, ICC, Flow Cyt, WBmore details
    Unsuitable for: IHC-P

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human lung, kidney, placenta and testis tissue lysates; HepG2 and Huh7 whole cell lysates. ICC: HepG2 cells. Flow cyt: HepG2 cells. IP: Huh7 whole cell lysate; Human lung tissue lysate.

  • General notes

    ab275394 is the carrier-free version of ab275390. 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

    ab275394 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

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C.

  • Storage buffer

    Constituent: 100% PBS

  • Carrier free

    Yes
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR23481-240

  • Isotype

    IgG

Images

  • Immunocytochemistry - Anti-COL18A1 antibody [EPR23481-240] - BSA and Azide free (ab275394)
    Immunocytochemistry - Anti-COL18A1 antibody [EPR23481-240] - BSA and Azide free (ab275394)

    This data was developed using ab275390, the same antibody clone in a different buffer formulation.

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling COL18A1 with ab275390 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HepG2 cells.  Negative control: U937 cell (PMID: 15451191). is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  • Flow Cytometry - Anti-COL18A1 antibody [EPR23481-240] - BSA and Azide free (ab275394)
    Flow Cytometry - Anti-COL18A1 antibody [EPR23481-240] - BSA and Azide free (ab275394)

    This data was developed using ab275390, the same antibody clone in a different buffer formulation.

    Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized U-937 (human histiocytic lymphoma monocyte, Left) / HepG2 (human hepatocellular carcinoma epithelial cell, Right) cells labelling COL18A1 with ab275390 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

    A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

    Negative control: U937 cell (PMID: 15451191).

  • Western blot - Anti-COL18A1 antibody [EPR23481-240] - BSA and Azide free (ab275394)
    Western blot - Anti-COL18A1 antibody [EPR23481-240] - BSA and Azide free (ab275394)
    All lanes : Anti-COL18A1 antibody [EPR23481-240] (ab275390) at 1/1000 dilution

    Lane 1 : HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate at 20 µg
    Lane 2 : Huh7 (human hepatocellar carcinoma epithelial cell), whole cell lysate at 40 µg
    Lane 3 : U937 (human histiocytic lymphoma monocyte), whole cell lysate at 40 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

    Predicted band size: 178 kDa
    Observed band size: 178,23,28 kDa
    why is the actual band size different from the predicted?



    This data was developed using ab275390, the same antibody clone in a different buffer formulation. 

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    The expression profile observed is consistent with what has been described in the literature (PMID: 11581192, 16465514, 15451191).

    Negative control: U937 (PMID:15451191).

    The lanes 1 of  blot were developed using a higher sensitivity ECL substrate.

    Exposure time: 92 seconds.

  • Immunoprecipitation - Anti-COL18A1 antibody [EPR23481-240] - BSA and Azide free (ab275394)
    Immunoprecipitation - Anti-COL18A1 antibody [EPR23481-240] - BSA and Azide free (ab275394)

    This data was developed using ab275390, the same antibody clone in a different buffer formulation.

    COL18A1 was immunoprecipitated from 0.35 mg Huh7 (human hepatocellular carcinoma epithelial cell) whole cell lysate with ab275390 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab275390 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: Huh7 (human hepatocellular carcinoma epithelial cell) whole cell lysate 10 ug

    Lane 2: ab275390 IP in Huh7 whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab275390 in Huh7 whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: Panel A: 32 seconds
    Panel B: 3 minutes

    Western blot was performed with the immuno-precipitate of ab275390, probed with ab275390 at 1/1000 dilution (Panel A) or ab207162 at 1/1000 dilution (Panel B). The 23- and 28-kDa fragments were recognized by ab207126, suggesting that these fragments are part of the C-terminal non-collagenous domain containing endostatin.

  • Immunoprecipitation - Anti-COL18A1 antibody [EPR23481-240] - BSA and Azide free (ab275394)
    Immunoprecipitation - Anti-COL18A1 antibody [EPR23481-240] - BSA and Azide free (ab275394)

    This data was developed using ab275390, the same antibody clone in a different buffer formulation.

    COL18A1 was immunoprecipitated from 0.35 mg Huh7 (human hepatocellular carcinoma epithelial cell) whole cell lysate with ab275390 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab275390 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: Huh7 (human hepatocellular carcinoma epithelial cell), whole cell lysate 10 ug

    Lane 2: ab275390 IP in Huh7 whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab275390 in Huh7 whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 101 seconds.

    23 kDa and 28 kDa fragments are part of the C-terminal non-collagenous domain containing endostatin.

  • Western blot - Anti-COL18A1 antibody [EPR23481-240] - BSA and Azide free (ab275394)
    Western blot - Anti-COL18A1 antibody [EPR23481-240] - BSA and Azide free (ab275394)
    All lanes : Anti-COL18A1 antibody [EPR23481-240] (ab275390) at 1/1000 dilution

    Lane 1 : Human lung tissue lysate
    Lane 2 : Human kidney tissue lysate
    Lane 3 : Human placenta tissue lysate
    Lane 4 : Human testis tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution

    Predicted band size: 178 kDa
    Observed band size: 23,28 kDa why is the actual band size different from the predicted?



    This data was developed using ab275390, the same antibody clone in a different buffer formulation. 

    locking and diluting buffer and concentration: 5% NFDM/TBST.

    The expression profile observed is consistent with what has been described in the literature (PMID: 11581192, 16465514, 15451191).

    Exposure time: Lane 1, 4: 3 minutes Lane 2, 3: 26 seconds.

  • Immunoprecipitation - Anti-COL18A1 antibody [EPR23481-240] - BSA and Azide free (ab275394)
    Immunoprecipitation - Anti-COL18A1 antibody [EPR23481-240] - BSA and Azide free (ab275394)

    This data was developed using ab275390, the same antibody clone in a different buffer formulation.

    COL18A1 was immunoprecipitated from 0.35 mg human lung tissue lysate with ab275390 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab275390 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: Human lung tissue lysate 10 ug

    Lane 2: ab275390 IP in Human lung tissue lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab275390 in Human lung tissue lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 7.75 seconds.

    23 kDa and 28 kDa fragments are part of the C-terminal non-collagenous domain containing endostatin.

  • Anti-COL18A1 antibody [EPR23481-240] - BSA and Azide free (ab275394)
    Anti-COL18A1 antibody [EPR23481-240] - BSA and Azide free (ab275394)

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