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 for FPN1 (NBP1-21502; 1:1000 dilution) and GAPDH (loading control). Image from verified customer review.

Immunocytochemistry/Immunofluorescence: Ferroportin/SLC40A1 Antibody [NBP1-21502] - HeLa cells were fixed and permeabilized for 10 minutes using -20C MeOH. The cells were incubated with anti-Ferroportin/SLC40A1 conjugated to Alexa Fluor 488 [NBp1-21502AF488] at 20ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunohistochemistry-Paraffin: Ferroportin/SLC40A1 Antibody [NBP1-21502] - IHC-P detection of FPN1 / SLC40A1 protein in murine small intestinal section using Ferroportin 1 antibody at a dilution of 1:200. The antibody primarily developed a membranous staining pattern in the intestinal epithelial cells.

Flow Cytometry: Ferroportin/SLC40A1 Antibody [NBP1-21502] - An intracellular stain was performed on HepG2 cells with Ferroportin/SLC40A1 Antibody NBP1-21502 and a matched isotype control. Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Western Blot: Ferroportin/SLC40A1 Antibody [NBP1-21502] - Human intestine lysate.

Western Blot: Ferroportin/SLC40A1 Antibody [NBP1-21502] - Bovine adult eye. Image was submitted via customer review.

Immunocytochemistry/Immunofluorescence: Ferroportin/SLC40A1 Antibody [NBP1-21502] - HepG2 cells were fixed and permeabilized for 10 minutes using -20C MeOH. The cells were incubated with anti-Ferroportin 1 at 5.0 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) [NB100-690] was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunohistochemistry-Paraffin: Ferroportin/SLC40A1 Antibody [NBP1-21502] - IHC-P detection of FPN1 / SLC40A1 protein in murine liver section using Ferroportin 1 antibody at a dilution of 1:200. The representative image shows intense staining in the cellular membranes, whereas, a relatively milder postivity was observed in the cytoplasm of hepatocytes.

Flow Cytometry: Ferroportin/SLC40A1 Antibody [NBP1-21502] - Histogram of Ferroportin staining of human spleen macrophages using Alexa Fluor 700 conjugated Ferroportin antibody (red, catalog# NBP1-21502AF700) and isotype control (grey) as a negative control. Image from verified customer review.

Flow Cytometry: Ferroportin/SLC40A1 Antibody [NBP1-21502] - Analysis using the PE conjugate of NBP1-21502. Staining of Ferroportin in human B lymphocytes in peripheral blood using PE conjugated anti-Ferroportin antibody. Image from verified customer review.

Flow Cytometry: Ferroportin/SLC40A1 Antibody [NBP1-21502] - An intracellular stain was performed on HepG2 cells with Ferroportin/SLC40A1 antibody NBP1-21502AF647 (blue) and a matched isotype control NBP2-24893AF647 (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Flow (Intracellular): Ferroportin/SLC40A1 Antibody [NBP1-21502] - An intracellular stain was performed on HepG2 cells with Ferroportin/SLC40A1 antibody NBP1-21502AF700 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 10 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 700.

• Reactivity: Hu, Mu, Po, Bv
• Applications: WB, Flow, Flow-IC, ICC/IF, IHC, IHC-P
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

Ferroportin/SLC40A1 Antibody Summary

• Immunogen: Synthetic peptide made to an internal portion of human Ferroportin 1 (within residues 250-300). [Swiss-Prot# Q9NP59]
• Localization: Cell membrane (high), Cytoplasm (low)
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: SLC40A1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1.0 ug/ml
  • Flow Cytometry 1:10
  • Immunocytochemistry/Immunofluorescence 1:500
  • Immunohistochemistry 1:200
  • Immunohistochemistry-Paraffin 1:200
  • Flow (Intracellular)
• Application Notes: This Ferroportin 1 antibody is useful for Immunocytochemistry/Immunofluorescence and Western blot, where a band is seen at ~62 kDa.
• Theoretical MW: 62 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.
• Control:

  Small Intestine Lysate (NB820-59255)

• Control Peptide:

  Ferroportin/SLC40A1 Peptide (NBP1-21502PEP)

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Immunogen affinity purified

Notes

Antibody tested in Flow Cytometry by verified customer through our Innovator's Reward Program.

Alternate Names for Ferroportin/SLC40A1 Antibody

• Ferroportin
• Ferroportin-1
• FPN1
• FPN1IREG1ferroportin 1
• HFE4
• HFE4ferroportin-1
• IREG1
• Iron-regulated transporter 1
• member 3
• MST079
• MTP1
• putative ferroportin 1 variant IIIB
• SLC11A3
• SLC11A3iron regulated gene 1
• SLC40A1
• solute carrier family 11 (proton-coupled divalent metal ion transporters)
• solute carrier family 40 (iron-regulated transporter), member 1
• solute carrier family 40 member 1

Background

Ferroportin 1 (FPN1 or SLC40A1) is an iron-regulated transporter (highly expressed in placenta, intestine, muscle, spleen etc.) that play key role in intestinal iron absorption as well as cellular iron release, and mediates iron efflux in the presence of ferroxidases hephaestin (HP) and/or ceruloplasmin (CP). FPN1 is implicated in the iron export from duodenal epithelial cell and also in transfer of iron between maternal and fetal circulation. FPN1 can be regulated transcriptionally, post-transcriptionally, through mRNA stability and post-translationally through protein turnover and its regulation is critical for iron homeostasis because alterations in the same may lead to either iron deficiency or iron overload. FPN1 transports iron in ferrous form whereas plasma transferrin only binds iron's ferric form, and 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 in the body 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. Defects in FPN1 have been linked to hemochromatosis type 4 (HFE4), an autosomal dominant iron-loading disorder.

Limitations

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.

Publications for Ferroportin/SLC40A1 Antibody (NBP1-21502)(38)

Publications using NBP1-21502

• Sajja RK, Kaisar MA, Vijay V et al. In Vitro Modulation of Redox and Metabolism Interplay at the Brain Vascular Endothelium: Genomic and Proteomic Profiles of Sulforaphane Activity Sci Rep Aug 23 2018 [PMID: 30139948] (ICC/IF, WB, Mouse)
• Ashok A, Karmakar S, Chandel R et al. Prion protein modulates iron transport in the anterior segment: Implications for ocular iron homeostasis and prion transmission Exp. Eye Res. May 31 2018 [PMID: 29859760] (ICC/IF, Human)
• Lim D, Kim KS, Jeong JH et al. The hepcidin-ferroportin axis controls the iron content of Salmonella-containing vacuoles in macrophages Nat Commun May 29 2018 [PMID: 29844422] (WB, Mouse)
• Broide E, Reifen R, Matalon S et al. Expression of Duodenal Iron Transporter Proteins in Diabetic Patients with and without Iron Deficiency Anemia. J Diabetes Res Mar 6 2018 [PMID: 29693022] (WB, Human)
• Puy V, Darwiche W, Trudel S et al. Predominant role of microglia in brain iron retention in Sanfilippo syndrome, a pediatric neurodegenerative disease. Glia Apr 6 2018 [PMID: 29624734] (WB, Mouse)
• McCarthy RC, SoSa JC, Gardeck AM et al. Inflammation-induced Iron Transport and Metabolism by Brain Microglia. J. Biol. Chem. Apr 2 2018 [PMID: 29610275] (Mouse)
• Zhabyeyev P, Das SK, Basu R et al. TIMP3 deficiency exacerbates iron-overload mediated cardiomyopathy and liver disease Am. J. Physiol. Heart Circ. Physiol. 2018 Jan 26 [PMID: 29373036] (WB, Mouse)
• Raggi C, Gammella E, Correnti M et al. Dysregulation of Iron Metabolism in Cholangiocarcinoma Stem-like Cells. Sci Rep 2017 Dec 15 [PMID: 29247214] (WB, Human)
• Zhang Fa-Li, Hou Hui-Min, Yin Zhi-Nan et al. Impairment of Hepcidin Upregulation by Lipopolysaccharide in the Interleukin-6 Knockout Mouse Brain. Frontiers in Molecular Neuroscience 2017 [PMID: 29163042] (WB, Mouse)
• Baba Y, Higa JK, Shimada BK et al. Protective Effects of the Mechanistic Target of Rapamycin against Excess Iron and Ferroptosis in Cardiomyocytes Am. J. Physiol. Heart Circ. Physiol. 2017 Nov 10 [PMID: 29127238] (Mouse)
• Neves JV, Ramos MF, Moreira AC et al. Hamp1 but not Hamp2 regulates ferroportin in fish with two functionally distinct hepcidin types Sci Rep. 2017 Nov 01 [PMID: 29093559] (Mouse)
• Zhang W, Lv T, Huang J, Ou X. Type 4B hereditary hemochromatosis associated with a novel mutation in the SLC40A1 gene: A case report and a review of the literature Medicine (Baltimore) 2017 Sep 01 [PMID: 28930842] (IHC, Human)
• Pasricha SR, Lim PJ, Duarte TL et al. Hepcidin is regulated by promoter-associated histone acetylation and HDAC3 Nat Commun 2017 Sep 01 [PMID: 28864822] (WB, Human)
• Dietrich P, Johnson IM, Alli S, Dragatsis I. Elimination of huntingtin in the adult mouse leads to progressive behavioral deficits, bilateral thalamic calcification, and altered brain iron homeostasis PLoS Genet. 2017 Jul 01 [PMID: 28715425] (WB, Mouse)
• Basuli D, Tesfay L, Deng Z et al. Iron addiction: a novel therapeutic target in ovarian cancer Oncogene 2017 Jul 20 [PMID: 28319068] (WB, Human)
• Fader KA, Nault R, Kirby MP et al. Convergence of hepcidin deficiency, systemic iron overloading, heme accumulation, and REV-ERBalpha/b activation in aryl hydrocarbon receptor-elicited hepatotoxicity. Toxicol. Appl. Pharmacol. 2017 Feb 16 [PMID: 28213091]
• Mertens C, Akam EA, Rehwald C et al. Intracellular Iron Chelation Modulates the Macrophage Iron Phenotype with Consequences on Tumor Progression. PLoS ONE. Nov 2 2016 [PMID: 27806101] (FLOW, Human)
• Zhang YT, Li FM, Guo YZ et al. (Z)-ligustilide increases ferroportin1 expression and ferritin content in ischemic SH-SY5Y cells. Eur. J. Pharmacol. Oct 24 2016 [PMID: 27789221] (WB)
• Gasperini L, Meneghetti E, Legname G, Benetti F. In Absence of the Cellular Prion Protein, Alterations in Copper Metabolism and Copper-Dependent Oxidase Activity Affect Iron Distribution. Front Neurosci. Oct 12 2016 [PMID: 27729845] (WB, Mouse)
• Asperti M, Stuemler T, Poli M et al. Heparanase Overexpression Reduces Hepcidin Expression, Affects Iron Homeostasis and Alters the Response to Inflammation. PLoS One 2016 Oct 06 [PMID: 27711215] (WB)
• Kim DK, Kim YH, Jung YS et al. Orphan nuclear receptor SHP regulates iron metabolism through inhibition of BMP6-mediated hepcidin expression. Sci Rep 2016 Sep 30 [PMID: 27688041] (WB)
• Marques O, Porto G, Rema A et al. Local iron homeostasis in the breast ductal carcinoma microenvironment BMC Cancer Mar 5 2016 [PMID: 26944411] (IHC-P, Human)
• Theurl M, Song D, Clark E et al. Mice with hepcidin-resistant ferroportin accumulate iron in the retina. FASEB J 2016 Feb [PMID: 26506980]
• Mori M, Ito F, Shi L et al. Ovarian endometriosis-associated stromal cells reveal persistently high affinity for iron. Redox Biol 2015 Dec [PMID: 26498255]
• Bose C, Megyesi JK, Shah SV et al. Evidence Suggesting a Role of Iron in a Mouse Model of Nephrogenic Systemic Fibrosis. PLoS One 2015 [PMID: 26305890] (WB, Mouse)
• Tesfay L, Clausen KA, Kim JW et al. Hepcidin regulation in prostate and its disruption in prostate cancer Cancer Res. 2015 Apr 09 [PMID: 25858146] (WB, Human)
• Gammella E, Diaz V, Recalcati S et al. Erythropoietin's inhibiting impact on hepcidin expression occurs indirectly. Am. J. Physiol. Regul. Integr. Comp. Physiol. 2014 Dec 17 [PMID: 25519735] (WB, Mouse)
• Katz O, Reifen R, Lerner A. B-Carotene can reverse dysregulation of iron protein in an in vitro model of inflammation. Immunol. Res. 2014 Nov 15 [PMID: 25398638] (WB, Human)
• Jeong SM, Lee J, Finley LW et al. SIRT3 regulates cellular iron metabolism and cancer growth by repressing iron regulatory protein 1. Oncogene. 2014 Jun 09 [PMID: 24909164] (WB, Mouse)
  
  Details:
  WB with Ferroportin 1 antibody on whole-cell lysates of immortalized WT and SIRT3 KO MEFs showing a decreased FPN1 protein expression in SIRT3 KO compared with WT cells (Figure 3e and Supplementary Figure S3f).
• Theurl M, Nairz M, Schroll A et al. Hepcidin as a predictive factor and therapeutic target in erythropoiesis-stimulating agent treatment for anemia of chronic disease in rats. Haematologica. 2014 Jun 03 [PMID: 24895335] (WB, IHC-P, Rat)
  
  Details:
  Formalin fixed and paraffin-wax embedded rat (female Lewis rats) spleen and duodenum tissue sections were immunostained with 1:200 dilutionof rabbit anti-Fp1 antibdoy (NBP1-21502) in PBS/1% BSA for 1h at RT followed by donkey anti rabbit Alexa 555 for immunofluorescence detection ( Supplemental Figure S4 - Splenic Fp1 and Supplemental Figure S5 - Duodenal Fp1 immunofluorescence in ACD rats treated with ESA and/or LDN-193189). Antibody also used for WB of FP1 in spleen (Figure 3A) and liver of rats (Supplemental Figure S3).
• Vercellotti Gregory M, Khan Fatima B, Nguyen Julia et al. H-ferritin ferroxidase induces cytoprotective pathways and inhibits microvascular stasis in transgenic sickle mice. Front Pharmacol. 2014 Apr 17 [PMID: 24860503] (WB, Mouse)
• Scheers NM, Almgren AB, Sandberg AS. Proposing a Caco-2/HepG2 cell model for in vitro iron absorption studies. The Journal of Nutritional Biochemistry 3/19/2014 [PMID: 24746839] (WB, Human)
• Kim DK, Jeong JH, Lee JM et al. Inverse agonist of estrogen-related receptor [gamma] controls Salmonella typhimurium infection by modulating host iron homeostasis. Nature Medicine 3/23/2014 [PMID: 24658075] (WB, Mouse)
• Scheers N, Sandberg AS. Iron Transport through Ferroportin Is Induced by Intracellular Ascorbate and Involves IRP2 and HIF2alpha. Nutrients 2014 Jan 3 [PMID: 24394537] (WB, Human)
• Diaz V, Gammella E, Recalcati S et al. Liver iron modulates hepcidin expression during chronically elevated erythropoiesis in mice. Hepatology 2013 Jun 6 [PMID: 23744538] (WB, Mouse)
• Choi JS, Koh IU, Lee HJ et al. Effects of excess dietary iron fat on glucose lipid metabolism. J Nutr Biochem 2013 May 2 [PMID: 23643521] (WB, Mouse)
• Bayeva M, Khechaduri A, Puig S et al. mTOR Regulates Cellular Iron Homeostasis through Tristetraprolin Cell Metab 2012 Oct 23 [PMID: 23102618] (WB, Mouse)
• Vanoaica L et al. Intestinal ferritin h is required for an accurate control of iron absorption. Cell Metab 12(3):273-82. 2010 [PMID: 20816093] (WB, Mouse)

Reviews for Ferroportin/SLC40A1 Antibody (NBP1-21502) (7)

Average Rating: 4.7

(Based on 7 reviews)

We have 7 reviews tested in 5 species: Human, Mouse, Bovine, Pig, bovine.

reviewed by: Anonymous WB Mouse 08/08/2018

Summary

• Application: Western Blot
• Sample Tested: Adult pancreas
• Species: Mouse
• Lot: F-1

reviewed by: Anonymous WB Pig 07/12/2018

Summary

• Application: Western Blot
• Sample Tested: duodenal mucosa
• Species: Pig
• Lot: F-1

reviewed by: Anonymous IHC-P Human 04/04/2018

Summary

• Application: Immunohistochemistry-Paraffin
• Sample Tested: Adult eye and muscle tissue (labeled E and M, respectively)
• Species: Human
• Lot: E-2

Comments

• Comments: 1:100

reviewed by: Anonymous WB bovine 04/04/2018

Summary

• Application: Western Blot
• Sample Tested: Adult eye
• Species: bovine
• Lot: E-2

Comments

• Comments: 1:1000

reviewed by: Anonymous IHC-P Bovine 10/24/2017

Summary

• Application: Immunohistochemistry-Paraffin
• Sample Tested: eye
• Species: Bovine
• Lot: E-2

Comments

• Comments: use at 1:100

reviewed by: Anonymous IHC Mouse 06/27/2017

Summary

• Application: Immunohistochemistry
• Sample Tested: Mouse brain
• Species: Mouse

reviewed by: Ajay Ashok WB Human 06/13/2017

Summary

• Application: Western Blot
• Sample Tested: Human microglia cell lysate
• Species: Human

FAQs for Ferroportin/SLC40A1 Antibody (NBP1-21502). (Showing 1 - 3 of 3 FAQs).

1. 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.
2. 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).
3. 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 our Ferroportin 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.

Bioinformatics

• Gene Symbol: SLC40A1
• Entrez:
  • 30061(Human)
  • 53945(Mouse)
  • 170840(Rat)
• Uniprot:
  • Q9NP59()