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    products/primary-antibodies/lipocalin-2--ngal-antibody-epr5084-ab125075.pdf

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Immunology Adaptive Immunity T Cells Non-CD
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Lipocalin-2 / NGAL antibody [EPR5084] (ab125075)

  • Datasheet
  • SDS
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Western blot - Anti-Lipocalin-2 / NGAL antibody [EPR5084] (ab125075)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipocalin-2 / NGAL antibody [EPR5084] (ab125075)
  • Western blot - Anti-Lipocalin-2 / NGAL antibody [EPR5084] (ab125075)
  • Immunocytochemistry/ Immunofluorescence - Anti-Lipocalin-2 / NGAL antibody [EPR5084] (ab125075)
  • Anti-Lipocalin-2 / NGAL antibody [EPR5084] (ab125075)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5084] to Lipocalin-2 / NGAL
  • Suitable for: WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

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Overview

  • Product name

    Anti-Lipocalin-2 / NGAL antibody [EPR5084]
    See all Lipocalin-2 / NGAL primary antibodies
  • Description

    Rabbit monoclonal [EPR5084] to Lipocalin-2 / NGAL
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore details
    Unsuitable for: Flow Cyt or IP
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Lipocalin-2/ NGAL aa 50-150. The exact sequence is proprietary.
    (Peptide available as ab187960)

  • Positive control

    • WB: Human fetal kidney lysate and SW480 cell lysate. IHC-P: human kidney tissue
  • 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.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR5084
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Adaptive Immunity
    • T Cells
    • Non-CD
    • Cardiovascular
    • Lipids / Lipoproteins
    • Fatty Acids
    • Binding proteins
    • Signal Transduction
    • Metabolism
    • Lipid metabolism
    • Cancer
    • Tumor biomarkers
    • Other
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Fatty acids
    • Metabolism
    • Types of disease
    • Cancer
    • Metabolism
    • Pathways and Processes
    • Redox metabolism
    • Fatty acid oxidation

Associated products

  • Alternative Versions

    • Anti-Lipocalin-2 / NGAL antibody [EPR5084] - BSA and Azide free (ab187969)
    • Alexa Fluor® 647 Anti-Lipocalin-2 / NGAL antibody [EPR5084] (ab206267)
  • Immunizing Peptide (Blocking)

    • Lipocalin-2 / NGAL peptide (ab187960)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human LCN2 knockout SW480 cell line (ab270486)
  • KO cell lysates

    • Human LCN2 knockout SW480 cell lysate (ab270509)
  • Positive Controls

    • SW480 whole cell lysate (ab3957)
  • Recombinant Protein

    • Recombinant Mouse Lipocalin-2 / NGAL protein (ab95008)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab125075 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
WB
1/1000 - 1/10000. Predicted molecular weight: 23 kDa.Can be blocked with Lipocalin-2 / NGAL peptide (ab187960).
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Antigen retrieval is recommended.

ICC/IF
1/100 - 1/250.
Notes
WB
1/1000 - 1/10000. Predicted molecular weight: 23 kDa.Can be blocked with Lipocalin-2 / NGAL peptide (ab187960).
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Antigen retrieval is recommended.

ICC/IF
1/100 - 1/250.
Application notes
Is unsuitable for Flow Cyt or IP.

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.
  • Target information above from: UniProt accession P80188 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3934 Human
    • Omim: 600181 Human
    • SwissProt: P80188 Human
    • Unigene: 204238 Human
    • 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 [EPR5084] (ab125075)
      Western blot - Anti-Lipocalin-2 / NGAL antibody [EPR5084] (ab125075)
      All lanes : Anti-Lipocalin-2 / NGAL antibody [EPR5084] (ab125075) 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 (Human lung carcinoma cell line) whole 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: 23 kDa
      Observed band size: 23 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab125075 observed at 23 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

      ab125075 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 ab125075 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 [EPR5084] (ab125075)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipocalin-2 / NGAL antibody [EPR5084] (ab125075)
      Immunohistochemical analysis of paraffin-embedded human kidney tissue sections labeling Lipocalin-2 / NGAL with purified ab125075 at 1/100 dilution. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Sections were counterstained with Hematoxylin.
      Antigen retrieval was heat mediated antigen retrieval using citrate buffer, pH 6.0).
    • Western blot - Anti-Lipocalin-2 / NGAL antibody [EPR5084] (ab125075)
      Western blot - Anti-Lipocalin-2 / NGAL antibody [EPR5084] (ab125075)
      All lanes : Anti-Lipocalin-2 / NGAL antibody [EPR5084] (ab125075) at 1/1000 dilution

      Lane 1 : Human fetal kidney lysate
      Lane 2 : SW480 (Human colorectal adenocarcinoma cell line) cell lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 23 kDa

    • Immunocytochemistry/ Immunofluorescence - Anti-Lipocalin-2 / NGAL antibody [EPR5084] (ab125075)
      Immunocytochemistry/ Immunofluorescence - Anti-Lipocalin-2 / NGAL antibody [EPR5084] (ab125075)
      Immunocytochemistry analysis of SW480 (human colorectal adenocarcinoma epithelial cell) labeling Lipocalin-2 / NGAL with purified ab125075 at 1/250 dilution (10 µg/ml). Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.9 µg/ml) was used as counterstain. Nuclei were stained blue with DAPI.
      Negative control: PBS instead of the primary antibody.
    • Anti-Lipocalin-2 / NGAL antibody [EPR5084] (ab125075)
      Anti-Lipocalin-2 / NGAL antibody [EPR5084] (ab125075)

    Protocols

    • Western blot protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (5)

    Publishing research using ab125075? Please let us know so that we can cite the reference in this datasheet.

    ab125075 has been referenced in 5 publications.

    • Uchimura K  et al. Human Pluripotent Stem Cell-Derived Kidney Organoids with Improved Collecting Duct Maturation and Injury Modeling. Cell Rep 33:108514 (2020). PubMed: 33326782
    • Lu Y  et al. CXCL1-LCN2 paracrine axis promotes progression of prostate cancer via the Src activation and epithelial-mesenchymal transition. Cell Commun Signal 17:118 (2019). PubMed: 31500632
    • Molina L  et al. Hepatocyte-Derived Lipocalin 2 Is a Potential Serum Biomarker Reflecting Tumor Burden in Hepatoblastoma. Am J Pathol 188:1895-1909 (2018). PubMed: 29920228
    • Ku AT  et al. TCF7L1 promotes skin tumorigenesis independently of ß-catenin through induction of LCN2. Elife 6:N/A (2017). WB . PubMed: 28467300
    • Wei T  et al. Elevated Expression of Matrix Metalloproteinase-9 not Matrix Metalloproteinase-2 Contributes to Progression of Extracranial Arteriovenous Malformation. Sci Rep 6:24378 (2016). WB . PubMed: 27075045

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