LCN2 Antibody - #DF6816

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Product: LCN2 Antibody
Catalog: DF6816
Description: Rabbit polyclonal antibody to LCN2
Application: WB IHC
Reactivity: Human, Mouse, Rat
Mol.Wt.: 23kDa; 23kD(Calculated).
Uniprot: P80188
RRID: AB_2838777

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Related Downloads
MS Report
HPLC Report
MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Clonality:
Polyclonal
Specificity:
LCN2 Antibody detects endogenous levels of total LCN2.
RRID:
AB_2838777
Cite Format: Affinity Biosciences Cat# DF6816, RRID:AB_2838777.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

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

Immunogens

Immunogen:
Uniprot:

P80188(NGAL_HUMAN) >>Visit HPA database.

Gene(ID):
LCN2(3934)
Expression:
P80188 NGAL_HUMAN:

Detected in neutrophils (at protein level) (PubMed:7683678, PubMed:8298140). 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.

Description:
Lipocalin-2, a member of the lipocalin family of proteins, was originally identified as a gelatinase-associated component of neutrophil secretory granules (1). Lipocalin-2 is involved in innate immunity, iron homeostasis, and apoptosis. Lipocalin-2 limits bacterial growth by binding to bacterial siderophores and sequestering iron (2-4). In mammalian cells, iron-loaded lipocalin-2 binds to its receptor, 24p3R, and is internalized, thereby releasing iron and increasing the intracellular iron concentration (5). Conversely, iron-free lipocalin-2 promotes apoptosis (5). Lipocalin-2 is also expressed in adipose tissue and promotes insulin resistance in cultured mouse adipocytes (6).
Sequence:
MPLGLLWLGLALLGALHAQAQDSTSDLIPAPPLSKVPLQQNFQDNQFQGKWYVVGLAGNAILREDKDPQKMYATIYELKEDKSYNVTSVLFRKKKCDYWIRTFVPGCQPGEFTLGNIKSYPGLTSYLVRVVSTNYNQHAMVFFKKVSQNREYFKITLYGRTKELTSELKENFIRFSKSLGLPENHIVFPVPIDQCIDG

PTMs - P80188 As Substrate

Site PTM Type Enzyme
Y72 Phosphorylation
Y76 Phosphorylation
N85 N-Glycosylation
Y126 Phosphorylation
S132 Phosphorylation
T133 Phosphorylation

Research Backgrounds

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 (By similarity). Involved in innate immunity; limits bacterial proliferation by sequestering iron bound to microbial siderophores, such as enterobactin. Can also bind siderophores from M.tuberculosis.

Subcellular Location:

Secreted. Cytoplasmic granule lumen. Cytoplasmic vesicle lumen.
Note: Upon binding to the SLC22A17 (24p3R) receptor, it is internalized (By similarity). Releases the bound iron in the acidic lumen of cytoplasmic vesicles (PubMed:12453413, PubMed:20581821).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Detected in neutrophils (at protein level). 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.

Subunit Structure:

Monomer. Homodimer; disulfide-linked. Heterodimer; disulfide-linked with MMP9.

Family&Domains:

Belongs to the calycin superfamily. Lipocalin family.

Research Fields

· Organismal Systems > Immune system > IL-17 signaling pathway.   (View pathway)

References

1). Myoglobin promotes macrophage polarization to M1 type and pyroptosis via the RIG-I/Caspase1/GSDMD signaling pathway in CS-AKI. Cell Death Discovery (PubMed: 35228524) [IF=7.0]

Application: IHC    Species: Mice    Sample: renal tissues

Fig. 7 DMF alleviated renal injury in CS-AKI mice. a Schematic diagram of experimental design of DMF treatment for CS-AKI mice. b, c qPCR analyses KIM-1 and NGAL mRNA level in the renal tissues of four groups (NC, CS, CS + DMF, DMF). d–g Concentration of biochemical indicators CK, myoglobin, SCr and BUN in serum. h Evaluation of the therapeutic effect of DMF on the kidney tissue of CS-AKI mice by HE staining (original magnification: 200×, scale bar: 100 μm). i IHC staining analyses the expression of NGAL in kidney tissues (original magnification: 200×, scale bar: 100 μm). For statistical analysis, one-way ANOVA followed by Tukey’s method for multiple comparisons used in (b–g). Data are expressed as mean ± SD, n = 3 per group. **P < 0.01, ***P < 0.001.
2). Roxadustat (FG-4592) protects against ischemia-induced acute kidney injury via improving CD73 and decreasing AIM2 inflammasome activation Get access Arrow. Nephrology Dialysis Transplantation (PubMed: 36413468) [IF=6.1]
3). Trans-cinnamaldehyde attenuates renal ischemia/reperfusion injury through suppressing inflammation via JNK/p38 MAPK signaling pathway. International Immunopharmacology (PubMed: 37011503) [IF=5.6]
4). Clinical significance of Interleukin 17 receptor E in diabetic nephropathy. International Immunopharmacology (PubMed: 37235960) [IF=5.6]

Application: IF/ICC    Species: Human    Sample: renal biopsy tissues

Fig. 4. Immunofluorescence double staining of IL-17RE and specific tubular/glomerular markers in renal biopsy tissues of patients with DN. IL-17RE was marked with FITC-conjugated secondary antibody; proximal tubular epithelial, distal tubular epithelial, and collecting duct cells were marked as AQP1, CD28k, and AQP3 with Cy3-conjugated secondary antibody, respectively; glomerular podocytes and endothelial and mesangial cells were marked as synaptopodin, CD31, and PDGFR-β with Cy3-conjugated secondary antibody, respectively. The white arrow indicates the positive staining area. NGAL, AQP1, CD28k, and AQP3: scale bar = 50 μm; synaptopodin, CD31, and PDGFR-β: scale bar = 25 μm. AQP, Aquaporin; CD28k, Calbindin D28k; PDGFR-β, platelet-derived growth factor receptors beta.

Application: IHC    Species: Human    Sample: renal biopsy tissues

Fig. 5. Pathological staining and protein expression levels in mouse kidney tissues. (A) H&E staining of the mouse kidneys. Scale bar = 50 μm. (B) PAS staining of mouse kidneys. Scale bar = 50 μm. (C) IHC assay of NGAL protein expression in the mouse kidneys. Scale bar = 50 μm. (D) IHC assay of IL-17RE protein expression in the mouse kidneys. Scale bar = 50 μm. The results represent the mean ± SD for six samples/group. *P < 0.05, ***P < 0.001 vs db/m. H&E, haematoxylin and eosin staining; PAS, periodic acid-Schiff staining; IHC, immunohistochemical staining.
5). Exosome‑encapsulated miR‑26a attenuates aldosterone‑induced tubulointerstitial fibrosis by inhibiting the CTGF/SMAD3 signaling pathway. International Journal of Molecular Medicine (PubMed: 36524378) [IF=5.4]

Application: WB    Species: Mice    Sample: kidneys

Figure 1 miR-26a expression is downregulated in the kidneys of ALD-induced mice. (A) Representative images of Masson's trichrome stained kidney tissues of mice in the sham and ALD groups. Scale bar, 50 µm. (B) RT-qPCR analysis of miR-26a expression levels in the kidney tissues of mice in the sham and ALD groups; U6 was used for normalization. (C) RT-qPCR analysis of collagen I, α-SMA and LCN2 mRNA expression levels in the kidney tissues of mice from the sham and ALD groups; β-actin was used for normalization. (D) Representative western blotting images and semi-quantitative analysis of E-cadherin, collagen I, α-SMA, CTGF and LCN2 protein expression levels in the kidney tissue of mice in the sham and ALD groups. (E) Immunohistochemical analysis of E-cadherin (green), α-SMA (red) and fibronectin (green) in the kidney tissue of mice in the sham and ALD groups; DAPI (blue) was used to stain the nuclei. Scale bar, 50 µm. Data are presented as the mean ± SD; n=5 mice/group; *P<0.05 vs. sham. α-SMA, α-smooth muscle actin2; CTGF, connective tissue growth factor; LCN2, lipocalin; RT-qPCR, reverse transcription-quantitative PCR.
6). Loganin Attenuates Septic Acute Renal Injury with the Participation of AKT and Nrf2/HO-1 Signaling Pathways. Drug Design Development and Therapy (PubMed: 33603340) [IF=4.8]
7). Calpain Inhibitor Calpeptin Alleviates Ischemia/Reperfusion-Induced Acute Kidney Injury via Suppressing AIM2 Inflammasome and Upregulating Klotho Protein. Frontiers in Medicine (PubMed: 35155498) [IF=3.9]

Application: WB    Species: Human    Sample: HK-2 cells

Figure 5 Calpeptin suppresses AIM2 and NLRP3 inflammasome signaling pathway, and increases Klotho protein expression in CoCl2-induced HK-2 cells hypoxia model. (A) Western blotting of Calpain 1, Calpain 2, Klotho, AIM2, NLRP3, pro-Caspase 1, cleaved-Caspase 1, ASC, IL-1β and LCN2 in the HK-2 cells. (B) Quantitative determination of Calpain 1, Calpain 2, Klotho, AIM2, NLRP3, cleaved-Caspase 1, IL-1β and LCN2. (C) The calpain activity of HK-2 cells was assessed by the relative fluorescence units (400/505 nm). (D) The cathepsin B activity of HK-2 cells was analyzed by the relative fluorescence units (400/505 nm). All data were presented as mean ± SEM (n = 3). NS, no significance; *p < 0.05 vs. control group; **p < 0.01 vs. control group; ***p < 0.001 vs. control group; #p < 0.05 vs. CoCl2 group; ##p < 0.01 vs. CoCl2 group; ###p < 0.001 vs. CoCl2 group. CP, calpeptin; RFU, relative fluorescence units.
8). Identification and analysis of a prognostic ferroptosis and iron-metabolism signature for esophageal squamous cell carcinoma. Journal of Cancer (PubMed: 35371305) [IF=3.9]

Application: IF/ICC    Species: Human    Sample: ESCC

Figure 4 Immunofluorescence of PRNP (green), SLC39A8 (red), LCN2 (red), ATP6V0A1 (green), SLC3A2 (red), and SLC39A14 (green) in ESCC samples and normal tissues.
9). N-acetylcysteine Ameliorates Vancomycin-induced Nephrotoxicity by Inhibiting Oxidative Stress and Apoptosis in the in vivo and in vitro Models. International Journal of Medical Sciences (PubMed: 35582415) [IF=3.6]
10). Antitumor effect of ginsenoside Rg3 on gallbladder cancer by inducing endoplasmic reticulum stress-mediated apoptosis in vitro and in vivo. Oncology Letters (PubMed: 30344724) [IF=2.9]

Application: WB    Species: human    Sample: GBC‑SD cells

Figure 1. |Rg3 activates ER stress in GBC‑SD cells. (A) GBC‑SD cells were treated with 1, 25, 50 and 100 µM Rg3 for 72 h, and proliferation was determined using a CCK‑8 kit. (B) Rg3 induced p‑eIF2α, ATF4 and Lcn2 expression in GBC‑SD cells. Relative protein expression level of (C) eIF2α, (D) p‑eIF2α, (E) ATF4 and (F) Lcn2 compared with the DMSO group. Relative protein expression was quantified by normalizing to the internal control β‑actin (n=3). *P<0.05, **P<0.01 vs. DMSO group. OD, optical density; ER, endoplasmic reticulum; eIF2α, eukaryotic translation‑initiation factor 2α; ATF4, activating transcription factor 4; p‑, phospho‑; DMSO, dimethylsulfoxide; Lcn2, lipocalin 2.

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