Recombinant 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
Related conjugates and formulations
Overview
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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 asab187960) -
Positive control
- WB: Human fetal kidney lysate and SW480 cell lysate. IHC-P: human kidney tissue
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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
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
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR5084 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Immunizing Peptide (Blocking)
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
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 |
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WB |
1/1000 - 1/10000. Predicted molecular weight: 23 kDa.Can be blocked with Lipocalin-2 / NGAL peptide (ab187960).
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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. |
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ICC/IF |
1/100 - 1/250.
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Notes |
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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. |
Target
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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
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Database links
- Entrez Gene: 3934 Human
- Omim: 600181 Human
- SwissProt: P80188 Human
- Unigene: 204238 Human
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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
see all
Images
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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 kDaLanes 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.
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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). -
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 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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (5)
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