Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Rat, Transgenic Mouse

Applications

Validated:

Immunohistochemistry, Western Blot

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Neutralization, Immunocytochemistry

Label

Unconjugated

Antibody Source

Monoclonal Rat IgG2A Clone # 220310
View all Lipocalin-2/NGAL Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human Lipocalin-2/NGAL
Gln21-Gly198
Accession # P80188

Specificity

Detects human Lipocalin-2/NGAL in direct ELISAs and Western blots. In direct ELISAs and Western blots, this antibody does not cross‑react with recombinant human (rh) Lipocalin-1 or rmLipocalin-2. This antibody also recognizes human Lipocalin‑2/MMP‑9 complexes in Western blots under non-reducing conditions.

Clonality

Monoclonal

Host

Rat

Isotype

IgG2A

Scientific Data Images for Human Lipocalin‑2/NGAL Antibody

Detection of Human Lipocalin‑2/NGAL antibody by Western Blot.

Detection of Human Lipocalin‑2/NGAL by Western Blot.

Western blot shows lysates of K562 human chronic myelogenous leukemia cell line. PVDF membrane was probed with 1 µg/mL of Rat Anti-Human Lipocalin-2/NGAL Monoclonal Antibody (Catalog # MAB1757) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for Lipocalin-2/NGAL at approximately 22 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Lipocalin-2/NGAL antibody in Human Pancreas by Immunohistochemistry (IHC-P).

Lipocalin‑2/NGAL in Human Pancreas.

Lipocalin-2/NGAL was detected in immersion fixed paraffin-embedded sections of human pancreas using Rat Anti-Human Lipocalin-2/NGAL Monoclonal Antibody (Catalog # MAB1757) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue). Specific staining was localized to plasma membrane of ductal cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human Lipocalin-2/NGAL antibody by Western Blot.

Detection of Human Lipocalin‑2/NGAL by Western Blot.

Western blot shows lysates of Capan-1 human pancreatic adenocarcinoma cell line, HT-29 human colon adenocarcinoma cell line, and human bone marrow tissue. PVDF membrane was probed with 0.2 µg/mL of Rat Anti-Human Lipocalin-2/NGAL Monoclonal Antibody (Catalog # MAB1757) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for Lipocalin-2/NGAL at approximately 22 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Lipocalin-2/NGAL by Western Blot

Detection of Lipocalin-2/NGAL by Western Blot

Silencing of LCN2 impairs cell cycle‐associated proteins. (A) The top proteins downregulated in LCN2‐silenced cells compared with control cells after RPPA proteomic analysis. (B) Gene set enrichment analysis of RPPA data identified pathways that are enriched or downregulated in control vs LCN2‐silenced SUM149 cells. (C) STRING interaction network of predicted active kinases based on enrichment of kinase substrates and protein interactions identified using KEA. The confidence of the interaction is reflected by the edge thickness. Based on node distribution analysis, four central proteins were identified (MAPK1, MAPK8, RPS6KB1, and MTOR). (D) Silencing of LCN2 in SUM149 cells reduced pMEK and pERK expression. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34342930), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Lipocalin-2/NGAL by Western Blot

Detection of Lipocalin-2/NGAL by Western Blot

LCN2 was highly expressed in tumors from patients with IBC. (A) High LCN2 expression was associated with shorter overall survival in a meta‐dataset of patients with non‐IBC. (B, C) LCN2 mRNA expression was higher in tumors from IBC patients vs non‐IBC patients in two independent breast cancer datasets [IBC World Consortium Dataset; GSE45582]. (D) LCN2 mRNA expression was higher in ER‐negative compared to ER+ samples IBC samples. (E) LCN2 mRNA expression was higher in more aggressive molecular subtypes, ERBB2+ and TNBC, compared to HR‐positive/HERBB2‐negative subtype. (F) LCN2‐high expression correlates with shorter overall survival in patients with IBC. (G) LCN2 mRNA expression was higher in IBC cell lines compared to non‐IBC cell lines. (H, I) LCN2 protein expression was higher in IBC cell lines compared to non‐IBC cell lines shown by (H) immunoblotting or (I) ELISA for secreted LCN2 in supernatants. Bar graphs indicate mean ± SEM from three independent experiments. graphpad prism software was used to obtain the P values, with Mann–Whitney tests used to compare two categories or one‐way analysis of variance to compare three or more categories. Black lines in each group (B–E, and G) indicate mean ± SD. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34342930), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human Lipocalin‑2/NGAL Antibody

Application
Recommended Usage

Immunohistochemistry

8-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human pancreas

Western Blot

0.2-1 µg/mL
Sample: K562 human chronic myelogenous leukemia cell line, Capan‑1 human pancreatic adenocarcinoma cell line, HT‑29 human colon adenocarcinoma cell line, and human bone marrow tissue

Reviewed Applications

Read 2 reviews rated 5 using MAB1757 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Lipocalin-2/NGAL

Members of Lipocalin family share a highly conserved fold with an eight-stranded antiparallel beta barrel, and act as a transporters, carrying small molecules to specific cells (1). Lipocalin-2, also known as Neutrophil Gelatinase-Associated Lipocalin (NGAL), was originally identified as a component of neutrophil granules (2). It is a 25 kDa protein existing in monomeric and homo- and heterodimeric forms, the latter as a dimer with human neutrophil gelatinases (MMP-9) (2). Its expression has been observed in most tissues normally exposed to microorganism, and its synthesis is induced in epithelial cells during inflammation (3). Lipocalin-2 has been implicated in a variety of processes including cell differentiation, tumorigenesis, and apoptosis (3-5). Studies indicate that Lipocalin-2 binds a bacterial catecholate sidropore bound to ferric ion such as enterobactin with a subnanomolar dissociation constant (Kd = 0.41 nM) (6). The bound ferric enterobactin complex breaks down slowly in a month into dihydroxybenzoyl serine and dihydroxybenzoic acid (DHBA). It also binds to a ferric DHBA complex with much less Kd values (7.9 nM) (6). Secretion of Lipocalin‑2 in immune cells increases by stimulation of Toll-like receptor as an acute phase response to infection. As a result, it acts as a potent bacteriostatic reagent by sequestering iron (7). Moreover, Lipocalin-2 can alter the invasive and metastatic behavior of Ras-transformed breast cancer cells in vitro and in vivo by reversing epithelial to mesenchymal transition inducing activity of Ras, through restoration of E-cadherin expression, via effects on the Ras-MAPK signaling pathway (8).

References

  1. Flower, D.R. et al. (1994) FEBS Lett. 354:7.
  2. Kjeldsen, L. et al. (1993) J. Biol. Chem. 268:10426.
  3. Kjeldsen L, et al. (2000) Biochim Biophys Acta. 1482:272.
  4. Devireddy, L.R. et al. (2001) Science 293:829.
  5. Yang, M.B. et al. (2002) Mol. Cell. 10:1045.
  6. Goetz, D.H. et al. (2002) Mol. Cell 10:1033.
  7. Flo, T.H. et al. (2004) Nature 432:917.
  8. Hanai, J. et al. (2005) J. Biol. Chem. 280:13641.

Long Name

Neutrophil Gelatinase-associated Lipocalin

Alternate Names

24p3, LCN2, Lipocalin2, MSFI, NGAL, Oncogene 24p3, p25, Siderocalin, Uterocalin

Entrez Gene IDs

3934 (Human); 16819 (Mouse); 170496 (Rat); 102137149 (Cynomolgus Monkey)

Gene Symbol

LCN2

UniProt

P80188

Additional Lipocalin-2/NGAL Products

Product Documents for Human Lipocalin‑2/NGAL Antibody

Certificate of Analysis

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Product Specific Notices for Human Lipocalin‑2/NGAL Antibody

For research use only

Citations for Human Lipocalin‑2/NGAL Antibody

Customer Reviews for Human Lipocalin‑2/NGAL Antibody (2)

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  • Human Lipocalin-2/NGAL Antibody
    Name: Anonymous
    Application: Immunohistochemistry
    Sample Tested: Lung tissue
    Species: Human
    Verified Customer | Posted 10/12/2021
    Formalin-fixed lung sections
    Human Lipocalin‑2/NGAL Antibody MAB1757
  • Human Lipocalin‑2/NGAL Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: A549 human lung carcinoma cell line
    Species: Human
    Verified Customer | Posted 02/24/2016
    Human Lipocalin‑2/NGAL Antibody MAB1757

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