Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-MCP3 antibody [EPR22649-155] (ab228979)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22649-155] to MCP3
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-MCP3 antibody [EPR22649-155]
    See all MCP3 primary antibodies

  • Description

    Rabbit monoclonal [EPR22649-155] to MCP3

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    IP
    Mouse
    WB
    Mouse
    Recombinant fragment
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: RAW264.7 (treated with 80 nM PMA), whole cell lysate; HEK-293T transfected with MCP3 expression vector containing a myc-His-tag®, whole lysate. IP: RAW264.7 (treated with 80 nM PMA for 24 hours, then with 100 ng/ml LPS treated for 6 hours with addition of 1ug/ml of Brefeldin A for the last 3 hours), cell 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

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR22649-155

  • Isotype

    IgG

  • Research areas

Images

  • Western blot - Anti-MCP3 antibody [EPR22649-155] (ab228979)
    Western blot - Anti-MCP3 antibody [EPR22649-155] (ab228979)
    All lanes : Anti-MCP3 antibody [EPR22649-155] (ab228979) at 1/1000 dilution

    Lane 1 : Wild-type RAW 264.7 untreated control cell lysate
    Lane 2 : Wild-type RAW 264.7 PMA treated (80 nM, 24 h) plus LPS treated (100 ng/ml, 6 h) and Brefeldin A (ab120299) treated (5 µg/ml, 5 h) cell lysate
    Lane 3 : CCL7/MCP3 knockout RAW 264.7 untreated cell lysate
    Lane 4 : CCL7/MCP3 knockout RAW 264.7 PMA treated (80 nM, 24 h) plus LPS treated (100 ng/ml, 6 h) and Brefeldin A (ab120299) treated (5 µg/ml, 5 h) cell lysate
    Lane 5 : J774A.1 Glucose treated (138.8 mMol/L, 8 h) plus Brefeldin A (ab120299) treated (5 µg/ml, 6 h) and Brefeldin A treated (5 µg/ml, 5 h) cell lysate
    Lane 6 : J774A.1 Glucose treated (5.6 mMol/L, 8 h) plus Brefeldin A (ab120299) treated (5 µg/ml, 6 h) and Brefeldin A treated (5 µg/ml, 5 h) cell lysate

    Lysates/proteins at 30 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 11 kDa
    Observed band size: 14 kDa
    why is the actual band size different from the predicted?



    Lanes 1 - 6: Merged signal (red and green). Green - ab228979 observed at 14 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

    ab228979 was shown to react with MCP3 in RAW 264.7 wild-type cells in Western blot with loss of signal observed in CCL7 knockout sample. Wild-type and CCL7 knockout RAW 264.7 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab228979 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.

  • Western blot - Anti-MCP3 antibody [EPR22649-155] (ab228979)
    Western blot - Anti-MCP3 antibody [EPR22649-155] (ab228979)
    All lanes : Anti-MCP3 antibody [EPR22649-155] (ab228979) at 1/1000 dilution

    Lane 1 : HEK-293T (human embryonic kidney) transfected with an empty vector (vector control), containing a myc-His-tag®, whole cell lysate
    Lane 2 : HEK-293T transfected with MCP3 expression vector containing a myc-His-tag®, whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 11 kDa
    Observed band size: 16-25 kDa why is the actual band size different from the predicted?



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

    Exposure time: 10 seconds.

  • Western blot - Anti-MCP3 antibody [EPR22649-155] (ab228979)
    Western blot - Anti-MCP3 antibody [EPR22649-155] (ab228979)
    All lanes : Anti-MCP3 antibody [EPR22649-155] (ab228979) at 1/1000 dilution

    Lane 1 : Untreated RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 40 µg
    Lane 2 : RAW264.7 (treated with 80 nM Phorbol-12-myristate-13-acetate (PMA) for 24 hours, then with 100 ng/ml lipopolysaccharides (LPS) treated for 6 hours with addition of 1ug/ml of Brefeldin A for the last 3 hours), whole cell lysate 40 ug.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 11 kDa
    Observed band size: 11 kDa



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

    Exposure time: 3 minutes.

  • Immunoprecipitation - Anti-MCP3 antibody [EPR22649-155] (ab228979)
    Immunoprecipitation - Anti-MCP3 antibody [EPR22649-155] (ab228979)

    MCP3 was immunoprecipitated from 0.35 mg RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) (treated with 80 nM Phorbol-12-myristate-13-acetate (PMA) for 24 hours, then with 100 ng/ml lipopolysaccharides (LPS) treated for 6 hours with addition of 1ug/ml of Brefeldin A for the last 3 hours), whole cell lysate with ab228979 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab228979 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.

    Lane 1: RAW264.7 (treated as above) whole cell lysate 10ug

    Lane 2: ab228979 IP in RAW264.7 (treated as above) whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab228979 in RAW264.7 (treated as above) whole cell lysate.

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

    Exposure time: 3 min.

     

  • Anti-MCP3 antibody [EPR22649-155] (ab228979)
    Anti-MCP3 antibody [EPR22649-155] (ab228979)

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