Western Blot: Ferroportin/SLC40A1 Antibody [NBP1-21502] - Western blot of human intestine lysate (Molecular weight: 62.5 KDa) using Ferroportin/SLC40A1 Antibody.
Western Blot: Ferroportin/SLC40A1 Antibody [NBP1-21502] - Duodenal mucosa samples from weanling pigs with iron replete or iron deficient status were probed using Ferroportin/SLC40A1 Antibody(NBP1-21502; 1:1000 ...read more
Flow Cytometry: Ferroportin/SLC40A1 Antibody [NBP1-21502] - An intracellular stain was performed on HepG2 cells with Ferroportin/SLC40A1 Antibody and a matched isotype control. Cells were fixed with 4% PFA and then ...read more
Western Blot: Ferroportin/SLC40A1 Antibody [NBP1-21502] - Mouse Ferroportin/SLC40A1, 5-aminolevulinic acid synthase (ALAS), and ferritin light chain (FLC) proteins are increased in sickle mice overexpressing human ...read more
Immunocytochemistry/ Immunofluorescence: Ferroportin/SLC40A1 Antibody [NBP1-21502] - HepG2 cells were fixed and permeabilized for 10 minutes using -20C MeOH. The cells were incubated with Ferroportin/SLC40A1 Antibody at ...read more
Immunohistochemistry-Paraffin: Ferroportin/SLC40A1 Antibody [NBP1-21502] - Detection of Ferroportin/SLC40A1 protein in murine small intestinal section using Ferroportin/SLC40A1 Antibody at a dilution of 1:200. The ...read more
Western Blot: Ferroportin/SLC40A1 Antibody [NBP1-21502] - Western blot of bovine adult eye using Ferroportin/SLC40A1 Antibody. Image from verified customer review.
Western Blot: Ferroportin/SLC40A1 Antibody [NBP1-21502] - HepG2 clones overexpressing heparanase showed a reduction of hepcidin expression and indices of iron loading. Two stable clones of HepG2 cells transfected with ...read more
Immunohistochemistry-Paraffin: Ferroportin/SLC40A1 Antibody [NBP1-21502] - Detection of Ferroportin/SLC40A1 protein in murine liver section using Ferroportin/SLC40A1 Antibody at a dilution of 1:200. The representative ...read more
Flow Cytometry: Ferroportin/SLC40A1 Antibody [NBP1-21502] - Analysis using the PE conjugate of Ferroportin/SLC40A1 Antibody. Staining of Ferroportin in human B lymphocytes in peripheral blood using PE conjugated ...read more
Flow Cytometry: Ferroportin/SLC40A1 Antibody [NBP1-21502] - An intracellular stain was performed on HepG2 cells with NBP1-21502AF647 (blue) and a matched isotype control (orange, NBP2-24893AF647). Cells were fixed with ...read more
62.5 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Rat reactivity reported in scientific literature (PMID: 24895335).
Packaging, Storage & Formulations
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
0.02% Sodium Azide
Immunogen affinity purified
Antibody tested in Flow Cytometry by verified customer through our Innovator's Reward Program.
Alternate Names for Ferroportin/SLC40A1 Antibody
iron regulated gene 1
Iron-regulated transporter 1
putative ferroportin 1 variant IIIB
SLC11A3iron regulated gene 1
solute carrier family 11 (proton-coupled divalent metal ion transporters)
solute carrier family 11 (proton-coupled divalent metal ion transporters), member 3
solute carrier family 40 (iron-regulated transporter), member 1
solute carrier family 40 member 1
Ferroportin is a 12-transmembrane domain protein, belonging to the major facilitator superfamily of transporters of small molecules, that is localized to the plasma membrane. Human Ferroportin has a theoretical molecular weight of 62.5 kDa. Ferroportin (FPN1 or SLC40A1) functions as an iron-regulated transporter (highly expressed in placenta, intestine, muscle, spleen, macrophages etc.) and is the receptor for the iron-regulatory hormone, hepcidin. In iron metabolism, FPN1 plays a key role in intestinal iron absorption as well as cellular iron release and mediates iron absorption in the presence of ferroxidases, hephaestin (HP) and/or ceruloplasmin (CP). FPN1 is implicated in iron export from duodenal epithelial cells and in the transfer of iron between maternal and fetal circulation. FPN1 transports iron in the ferrous form whereas plasma transferrin only binds iron's ferric form. Ferroxidases are key players in oxidizing iron transported by FPN1 and without the activity of ferroxidases, FPN1 is internalized followed by degradation. While other cell types utilize the circulating or GPI-linked multicopper ferroxidase CP for FPN1, intestinal cells utilize a membrane-bound HP, a paralog of CP that also show interaction with FPN1 (1).
FPN1 regulation is dependent on the cell type and involves transcriptional, posttranscriptional, and posttranslational mechanisms including hepcidin-mediated endocytosis and proteolysis. Hepcidin controls the concentration of FPN1 in the membrane, with hepcidin deficiency resulting in iron overload (high iron) and hepcidin excess leading to iron restriction and anemia (2). Ferroportin disease or hemochromatosis type 4 (HFE4) is associated with distinct FPN1 variants with either reduced FPN1 cell surface expression/iron export capacity or hepcidin resistance and iron overload (3, 4).
1. De Domenico I, Ward DM, Kaplan J. (2011) Hepcidin and ferroportin: the new players in iron metabolism. Semin Liver Dis. 31(3):272-9. PMID: 21901657
2. Drakesmith H, Nemeth E, Ganz T. (2015) Ironing out Ferroportin. Cell Metab. 22(5):777-87. PMID: 26437604
3. Pietrangelo A. (2017) Ferroportin disease: pathogenesis, diagnosis and treatment. Haematologica. 102(12):1972-1984. PMID: 29101207
4. Vlasveld LT, Janssen R, Bardou-Jacquet E, Venselaar H, Hamdi-Roze H, Drakesmith H, Swinkels DW. (2019) Twenty Years of Ferroportin Disease: A Review or An Update of Published Clinical, Biochemical, Molecular, and Functional Features. Pharmaceuticals (Basel). 12(3). pii: E132. PMID: 31505869
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
FAQs for Ferroportin/SLC40A1 Antibody (NBP1-21502). (Showing 1 - 5 of 5 FAQs).
I have some leftovers of this antibody from two years ago and tried it for immunofluorescence on mouse, which did work well. I may need more of this antibody. Some things surprise me. In the old datasheet it says that there is cross reactivity with mouse. It does not say so in the new data sheet here on the web site. However, it does say so for all the conjugated versions of the Ab. What is correct? Also, in the old data sheet it says that it is for Western blotting. Now immunofluorescence is also mentioned. Is this still the same antibody? I don't have a lot number, but we got it in September 2010.
NBP1-21502 is still the same antibody it was two years ago. Mouse was mistakenly removed during a website update and has since been re-added. This antibody still cross-reacts with mouse. In the past two years we have begun to validate more of our products for use in ICC and this is one that has had its uses updated since you last purchased.
Would it be possible to conjugate this antibody to PE or PerCP?
With regards to conjugating NBP1-21502SS (0.025 ml) to PE or PerCP, there are two possibilities: The first is that you could conjugate the antibody yourself, using easy-to-use kits. You would choose from Lightning-Link R-PE 3 x 0.01 mg, or Lightning-Link PerCP 3 x 0.01 mg. Because the buffer of the NBP1-21502 antibody is Tris-glycine, which is not compatible with the Lightning-Link conjugation chemistry as that targets amine groups, you would perform a buffer exchange first using the Antibody Concentration and Clean Up Kit. The lab has informed me that yes, they could conjugate your choice of PE or PerCP to this antibody for you. However, it would have to be the full-size antibody vial (0.1 ml) rather than the sample size (0.025 ml).
Is the epitope for NBP1-21502 extracellular or will it require permeabilization? Is it possible to get a sample size of one of the conjugated versions?
The immunogen used for this Ferroportin/SLC40A1 Antibody NBP1-21502 falls within a helical domain of the protein and some permeabilization is recommended for staining. Our lab uses 0.2% Tween-20 included in the blocking and diluent buffers to perform this permeabilization. As our directly conjugated antibodies are prepared per order, we unfortunately do not have sample sizes available.
I can’t get positive a result in IHC-P using NBP1-21502 (tissue: atherosclerosis hemorrhage plaque). PV-6001 and DAB is working. Can you make optimization suggestions on how to get this antibody to work?
Optimization could really make a difference as we have seen some staining with the same antibody on frozen section. The main thing would be to focus on antigen retrieval step and the buffer used. For the antigen retrieval method, we would suggest an increase to at least 10-20 minutes to the pressure cooker step of the HIER method; the duration of the retrieval step can greatly determine if the antigen will be exposed or not for the antigen binding site of the antibody to recognize the protein. Also, I the incubation time for peroxidase blocking time could be reduced from 30min to 15 min, as we have mentioned. Since this is a TMP, permeabilization reagents should be used for the experiment. You can review what we have available on our site in terms of our general protocols:HIER: https://www.novusbio.com/support/support-by-application/antigen-retrieval/protocol.htmlIHC-P: https://www.novusbio.com/support/support-by-application/immunohistochemistry-paraffin/protocol.html
We are having difficulty producing a clean blot, i.e. when we probe our western blots with the recommended concentration of 1.0 ug/ml, we are seeing multiple bands. The order of the molecular weight ladder we used is as followed: 250, 130, 100, 70, 55, 35, 25, 15, and 10 kDa. Are you able to indicate what band may be ferroportin.?
There is a western blot protocol specific to this antibody that can be found on the datasheet; it differs from your protocol quite a lot. I would give this protocol a try and see if you get better results.https://resources.novusbio.com/protocols/Western-Blot-Protocol-for-Ferroportin-1-Antibody-(NBP1-21502).pdfAlthough I am uncertain of your blocking method, we find that BSA and PVDF can give high non-specific background so the nitrocellulose and NFDM are recommend for this antibody. In addition, there seems to be no protein expression of the SLC40A1 in liver or spleen based on Human Protein Atlas.https://www.proteinatlas.org/ENSG00000138449-SLC40A1/tissueA good positive control might be bone marrow.
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Bio-Techne appreciates the critical role that you and our products and services play in research efforts to further scientific innovation and discovery. We are continually assessing our manufacturing and supplier capabilities during the COVID-19 situation and are implementing precautionary measures to ensure uninterrupted supply of products and services. Currently, and as we abide by local shelter in place orders across the world, we are fully operational and do not anticipate any material supply disruptions across our Bio-Techne brands and product lines. As the situation evolves, our goal is to utilize preventive measures to reduce the threat that COVID-19 poses to our ability to meet the needs of our customers globally.