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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Lipocalin-2 / NGAL antibody [EPR19912] (ab206427)

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Western blot - Anti-Lipocalin-2 / NGAL antibody [EPR19912] (ab206427)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipocalin-2 / NGAL antibody [EPR19912] (ab206427)
  • Western blot - Anti-Lipocalin-2 / NGAL antibody [EPR19912] (ab206427)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipocalin-2 / NGAL antibody [EPR19912] (ab206427)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipocalin-2 / NGAL antibody [EPR19912] (ab206427)
  • Anti-Lipocalin-2 / NGAL antibody [EPR19912] (ab206427)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19912] to Lipocalin-2 / NGAL
  • Suitable for: IHC-P, WB
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-Lipocalin-2 / NGAL antibody [EPR19912]
    See all Lipocalin-2 / NGAL primary antibodies

  • Description

    Rabbit monoclonal [EPR19912] to Lipocalin-2 / NGAL

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, WBmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: SW480 and A431 whole cell lysates; human ovary cancer, fetal spleen and colon cancer lysates; IHC-P: Human spleen, liver and ovary cancer tissues.

  • 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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab206427 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
IHC-P
1/8000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
1/1000. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).
Notes
IHC-P
1/8000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
1/1000. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).

Target

  • Function

    Iron-trafficking protein involved in multiple processes such as apoptosis, innate immunity and renal development. Binds iron through association with 2,5-dihydroxybenzoic acid (2,5-DHBA), a siderophore that shares structural similarities with bacterial enterobactin, and delivers or removes iron from the cell, depending on the context. Iron-bound form (holo-24p3) is internalized following binding to the SLC22A17 (24p3R) receptor, leading to release of iron and subsequent increase of intracellular iron concentration. In contrast, association of the iron-free form (apo-24p3) with the SLC22A17 (24p3R) receptor is followed by association with an intracellular siderophore, iron chelation and iron transfer to the extracellular medium, thereby reducing intracellular iron concentration. Involved in apoptosis due to interleukin-3 (IL3) deprivation: iron-loaded form increases intracellular iron concentration without promoting apoptosis, while iron-free form decreases intracellular iron levels, inducing expression of the proapoptotic protein BCL2L11/BIM, resulting in apoptosis. Involved in innate immunity, possibly by sequestrating iron, leading to limit bacterial growth.

  • Tissue specificity

    Expressed in bone marrow and in tissues that are prone to exposure to microorganism. High expression is found in bone marrow as well as in uterus, prostate, salivary gland, stomach, appendix, colon, trachea and lung. Not found in the small intestine or peripheral blood leukocytes.

  • Sequence similarities

    Belongs to the calycin superfamily. Lipocalin family.

  • Cellular localization

    Secreted. Upon binding to the SLC22A17 (24p3R) receptor, it is internalized.
  • Information by UniProt

  • Database links

    • Alternative names

      • 24p3 antibody
      • 25 kDa alpha 2 microglobulin related subunit of MMP9 antibody
      • 25 kDa alpha-2-microglobulin-related subunit of MMP-9 antibody
      • Alpha 2 microglobulin related protein antibody
      • HGNC:6526 antibody
      • HNL antibody
      • Lcn 2 antibody
      • Lcn2 antibody
      • Lipocalin-2 antibody
      • Migration stimulating factor inhibitor antibody
      • MSFI antibody
      • Neutrophil gelatinase associated lipocalin antibody
      • Neutrophil gelatinase associated lipocalin precursor antibody
      • Neutrophil gelatinase-associated lipocalin antibody
      • NGAL antibody
      • NGAL_HUMAN antibody
      • Oncogene 24p3 antibody
      • Oncogenic lipocalin 24p3 antibody
      • p25 antibody
      • Siderocalin antibody
      • siderocalin LCN2 antibody
      • SV40 induced 24P3 protein antibody
      • Uterocalin antibody
      see all

    Images

    • Western blot - Anti-Lipocalin-2 / NGAL antibody [EPR19912] (ab206427)
      Western blot - Anti-Lipocalin-2 / NGAL antibody [EPR19912] (ab206427)
      All lanes : Anti-Lipocalin-2 / NGAL antibody [EPR19912] (ab206427) at 1/1000 dilution

      Lane 1 : Wild-type SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate
      Lane 2 : LCN2 knockout SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate
      Lane 3 : A549 cell lysate
      Lane 4 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 22 kDa
      Observed band size: 25 kDa why is the actual band size different from the predicted?



      Lanes 1 - 4: Merged signal (red and green). Green - ab206427 observed at 25 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

      ab206427 was shown to react with Lipocalin-2 / NGAL in wild-type SW480 cells in Western blot with loss of signal observed in LCN2 knockout cell line ab270486 (knockout cell lysate ab270509). Wild-type SW480 and LCN2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab206427 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipocalin-2 / NGAL antibody [EPR19912] (ab206427)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipocalin-2 / NGAL antibody [EPR19912] (ab206427)

      Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling Lipocalin-2 / NGAL with ab206427 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Positive staining on neutrophils is observed. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Western blot - Anti-Lipocalin-2 / NGAL antibody [EPR19912] (ab206427)
      Western blot - Anti-Lipocalin-2 / NGAL antibody [EPR19912] (ab206427)
      All lanes : Anti-Lipocalin-2 / NGAL antibody [EPR19912] (ab206427) at 1/1000 dilution

      Lane 1 : A431 (Human epidermoid carcinoma cell line) whole cell lysate
      Lane 2 : Human ovary cancer lysate
      Lane 3 : Human fetal spleen lysate
      Lane 4 : Human colon cancer lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      Lane 1 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
      Lanes 2-4 : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/100000 dilution

      Predicted band size: 22 kDa
      Observed band size: 22 kDa



      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure time: Lane 1-3: 3 minutes; Lane 4: 30 seconds.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipocalin-2 / NGAL antibody [EPR19912] (ab206427)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipocalin-2 / NGAL antibody [EPR19912] (ab206427)

      Immunohistochemical analysis of paraffin-embedded human liver tissue labeling Lipocalin-2 / NGAL with ab206427 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Positive staining on neutrophils and negative on hepatocytes. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipocalin-2 / NGAL antibody [EPR19912] (ab206427)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipocalin-2 / NGAL antibody [EPR19912] (ab206427)

      Immunohistochemical analysis of paraffin-embedded human ovary cancer tissue labeling Lipocalin-2 / NGAL with ab206427 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human ovary cancer is observed. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Anti-Lipocalin-2 / NGAL antibody [EPR19912] (ab206427)
      Anti-Lipocalin-2 / NGAL antibody [EPR19912] (ab206427)

    References (0)

    Publishing research using ab206427? Please let us know so that we can cite the reference in this datasheet.

    ab206427 has not yet been referenced specifically in any publications.

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