Recombinant Anti-Lipocalin-2 / NGAL antibody [EPR19912] (ab206427)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19912] to Lipocalin-2 / NGAL
- Suitable for: IHC-P, WB
- Knockout validated
- Reacts with: Human
Overview
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Product name
Anti-Lipocalin-2 / NGAL antibody [EPR19912]
See all Lipocalin-2 / NGAL primary antibodies -
Description
Rabbit monoclonal [EPR19912] to Lipocalin-2 / NGAL -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WBmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: SW480 and A431 whole cell lysates; human ovary cancer, fetal spleen and colon cancer lysates; IHC-P: Human spleen, liver and ovary cancer tissues.
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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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19912 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lysates
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab206427 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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IHC-P |
1/8000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
1/1000. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).
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Notes |
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IHC-P
1/8000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa). |
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 [EPR19912] (ab206427) 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: 22 kDa
Observed band size: 25 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab206427 observed at 25 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab206427 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 ab206427 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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Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling Lipocalin-2 / NGAL with ab206427 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Positive staining on neutrophils is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-Lipocalin-2 / NGAL antibody [EPR19912] (ab206427) at 1/1000 dilution
Lane 1 : A431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 2 : Human ovary cancer lysate
Lane 3 : Human fetal spleen lysate
Lane 4 : Human colon cancer lysate
Lysates/proteins at 10 µg per lane.
Secondary
Lane 1 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lanes 2-4 : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/100000 dilution
Predicted band size: 22 kDa
Observed band size: 22 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1-3: 3 minutes; Lane 4: 30 seconds.
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Immunohistochemical analysis of paraffin-embedded human liver tissue labeling Lipocalin-2 / NGAL with ab206427 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Positive staining on neutrophils and negative on hepatocytes. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded human ovary cancer tissue labeling Lipocalin-2 / NGAL with ab206427 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human ovary cancer is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (1)
ab206427 has been referenced in 1 publication.
- Chen W et al. Lipocalin-2 Exacerbates Lupus Nephritis by Promoting Th1 Cell Differentiation. J Am Soc Nephrol 31:2263-2277 (2020). PubMed: 32646856