Anti-Lipocalin-2 / NGAL antibody (ab63929)

Rabbit polyclonal Lipocalin-2 / NGAL antibody. Validated in WB, ICC/IF and tested in Mouse, Rat, Human. Cited in 18 publication(s). Independently reviewed in 4 review(s).

Overview

  • Product name
    Anti-Lipocalin-2 / NGAL antibody
    See all Lipocalin-2 / NGAL primary antibodies
  • Description
    Rabbit polyclonal to Lipocalin-2 / NGAL
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Mouse Lipocalin-2/ NGAL. Residues surrounding amino acid 40.

  • Positive control
    • WB: Jurkat cell lysate. Recombinant mouse Lipocalin-2/ NGAL protein. ICC/IF: NIH/3T3 cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab63929 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 Use a concentration of 1 - 4 µg/ml. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa).
ICC/IF 1/100.

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

  • Anti-Lipocalin-2 / NGAL antibody (ab63929) at 1/1000 dilution (12 hours at 4degC) + Rat kidney whole tissue lysate at 10 µg

    Secondary
    Goat anti-mouse IgG-HRP at 1/10000 dilution

    Developed using the ECL technique.

    Performed under non-reducing conditions.

    Predicted band size: 23 kDa


    Exposure time: 10 seconds


    Blocking: 5% milk for 1 hour at 20°C.

  • All lanes : Anti-Lipocalin-2 / NGAL antibody (ab63929) at 1.5 µg/ml

    Lane 1 : Uninduced Jurkat cell lysate
    Lane 2 : Induced Jurkat cell lysate

    Lysates/proteins at 35 µg per lane.

    Predicted band size: 23 kDa
    Observed band size: 23 kDa
    Additional bands at: 70 kDa (possible non-specific binding)

  • All lanes : Anti-Lipocalin-2 / NGAL antibody (ab63929) at 2 µg/ml (12 hours at 4deg)

    Lanes 1 & 4 : Healthy mouse urine treated with CLB (NP-40 based lysis buffer with protease inhibitors)
    Lanes 2-3 : Acute kidney injury mouse urine treated with CLB (NP-40 based lysis buffer with protease inhibitors)

    Lysates/proteins at 5 µg per lane.

    Secondary
    All lanes : Donkey anti rabbit HRP at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 23 kDa


    Exposure time: 1 second


    Diluent: TBS - 0.1 % Tween- 5% milk

  • All lanes : Anti-Lipocalin-2 / NGAL antibody (ab63929) at 4 µg/ml

    Lane 1 : Recombinant mouse Lipocalin-2/ NGAL protein, 2 ng
    Lane 2 : Recombinant mouse Lipocalin-2/ NGAL protein, 10 ng
    Lane 3 : Recombinant mouse Lipocalin-2/ NGAL protein, 50 ng

    Predicted band size: 23 kDa

  • ab63929 staining Lipocalin-2 / NGAL in the RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) cell line by ICC/IF (Immunocytochemistry/immunofluorescence).

    Cells were fixed with paraformaldehyde, permeabilized with Triton 0.1% + 2% BSA in PBS and blocked with 2% BSA for 30 minutes at 24°C. Samples were incubated with primary antibody (1/100 in PBS + 2% BSA) for 16 hours at 4°C. An Alexa Fluor®488-conjugated Goat anti-rabbit polyclona l(1/1000) was used as the secondary antibody.

    See Abreview

  • NIH/3T3 (Mouse embryo fibroblast cell line) cells labeling Lipocalin-2 / NGAL using ab63929 (red) in ICC/IF.

References

This product has been referenced in:
  • Festa BP  et al. Impaired autophagy bridges lysosomal storage disease and epithelial dysfunction in the kidney. Nat Commun 9:161 (2018). Read more (PubMed: 29323117) »
  • Betten R  et al. Tonicity inversely modulates lipocalin-2 (Lcn2/24p3/NGAL) receptor (SLC22A17) and Lcn2 expression via Wnt/ß-catenin signaling in renal inner medullary collecting duct cells: implications for cell fate and bacterial infection. Cell Commun Signal 16:74 (2018). Read more (PubMed: 30404645) »
See all 18 Publications for this product

Customer reviews and Q&As

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1-4 of 4 Abreviews

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (kidney)
Antigen retrieval step
None
Permeabilization
No
Specification
kidney
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Feb 12 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - whole (kidney)
Gel Running Conditions
Non-reduced Denaturing (12%)
Loading amount
10 µg
Specification
kidney
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Jan 29 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (RAW264.7)
Specification
RAW264.7
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton + 2% BSA in PBS
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 24°C

Dr. Mahesh Shivananjappa

Verified customer

Submitted Apr 16 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - other (urine)
Loading amount
5 µg
Specification
urine
Treatment
CLB (NP-40 based lysis buffer with protease inhibitors)
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Nov 23 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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