Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-COL18A1 antibody [EPR23481-240] (ab275390)

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
  • Suitable for: Flow Cyt, WB, ICC, IP
  • Reacts with: Human

Overview

  • Product name

    Anti-COL18A1 antibody [EPR23481-240]
    See all COL18A1 primary antibodies

  • Description

    Rabbit monoclonal [EPR23481-240] to COL18A1

  • Host species

    Rabbit

  • Specificity

    This antibody does not recognise Endostatin.

    This antibody ab275390 [EPR23481-240]] is a distinct clone from ab275746 [EPR23481-21].

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • 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

    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.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.

  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59.94% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR23481-240

  • Isotype

    IgG

Images

  • Immunocytochemistry - Anti-COL18A1 antibody [EPR23481-240] (ab275390)
    Immunocytochemistry - Anti-COL18A1 antibody [EPR23481-240] (ab275390)

    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] (ab275390)
    Flow Cytometry - Anti-COL18A1 antibody [EPR23481-240] (ab275390)

    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] (ab275390)
    Western blot - Anti-COL18A1 antibody [EPR23481-240] (ab275390)
    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?



    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] (ab275390)
    Immunoprecipitation - Anti-COL18A1 antibody [EPR23481-240] (ab275390)

    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] (ab275390)
    Immunoprecipitation - Anti-COL18A1 antibody [EPR23481-240] (ab275390)

    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] (ab275390)
    Western blot - Anti-COL18A1 antibody [EPR23481-240] (ab275390)
    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?



    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).

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

     

  • Immunoprecipitation - Anti-COL18A1 antibody [EPR23481-240] (ab275390)
    Immunoprecipitation - Anti-COL18A1 antibody [EPR23481-240] (ab275390)

    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] (ab275390)
    Anti-COL18A1 antibody [EPR23481-240] (ab275390)

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