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Purified anti-FBXL5 Antibody

Pricing & Availability
Clone
10F4H9D12 (See other available formats)
Regulatory Status
RUO
Other Names
F-BOX/LLR-repeat protein 5, FBL5
Isotype
Mouse IgG2a
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Product Citations
publications
10F4H9D12_Purified_FBXL5_Antibody_1_072418
Total cell lysates (30 µg protein) from HeLa cells treated with 100 µM desferal (Dfo) overnight, untreated HeLa cells (-), or HeLa cells treated with 100 µg/mL FeCl3 (Fe) for 2 hours were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to nitrocellulose, and probed with the 0.5 µg/mL (1:1000 dilution) of purified anti-FBXL5 antibody (clone 10F4H9D12). Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse-IgG (Cat. No. 405301) at a 1:3000 dilution. Equal protein loading was confirmed using Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 649203) at a 1:2000 dilution. Lane M: MW ladder; n.s band: non-specific band.
  • 10F4H9D12_Purified_FBXL5_Antibody_1_072418
    Total cell lysates (30 µg protein) from HeLa cells treated with 100 µM desferal (Dfo) overnight, untreated HeLa cells (-), or HeLa cells treated with 100 µg/mL FeCl3 (Fe) for 2 hours were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to nitrocellulose, and probed with the 0.5 µg/mL (1:1000 dilution) of purified anti-FBXL5 antibody (clone 10F4H9D12). Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse-IgG (Cat. No. 405301) at a 1:3000 dilution. Equal protein loading was confirmed using Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 649203) at a 1:2000 dilution. Lane M: MW ladder; n.s band: non-specific band.
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672603 25 µg $101
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672604 100 µg $253
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Description

In metazoans, iron homeostasis is mediated at the systemic and cellular levels by the RNA-binding iron regulatory proteins IRP1 and IRP2.Under conditions where cellular iron is limiting, IRP1 and IRP2 stabilize the mRNA of genes involved in iron acquisition and mobilization and translationally repress genes involved in iron storage and export.  Under iron replete conditions, both IRPs are inactivated: IRP1 acquires an Fe-S cluster that inhibits RNA-binding activity, while IRP2 is targeted for proteasomal-mediated degradation by the E3 ubiquitin ligase FBXL5. Under iron-replete conditions Fe-S cluster biogenesis is disrupted, FBXL5 will also target IRP1 for degradation.

FBXL5, or F-box and leucine-rich repeat protein 5 (FBXL5), is the E3 ubiquitin ligase component of the SCFFBXL5 complex. The complex contains SKP1, Cullin-1, RBX1, and FBXL5. When cytosolic iron is replete, FBXL5 recruits IRP2 to the SCF complex and promotes its ubiquitination and subsequent degradation. FBXL5 contains a hemerythrin-like domain that senses oxygen and iron availability. This sensing mechanism is linked to the stability of the FBXL5 protein itself: when cytosolic iron is available, iron binds to the hemerythrin domain and promotes the stability of the FBXL5 protein. When iron is deficient, the FBXL5 protein lacks the iron ligand, resulting in its degradation and activation of the IRP-mediated iron starvation response.

Studies have also shown that FBXL5 can ubiquitinate and target the SNAIL1 transcription factor and disruption of this process promotes epithelialto mesenchymal transition. 

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Recombinant human FBXL5
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
Recommended Usage

Each lot of this antibody is quality control tested by Western blotting. For Western blotting, the suggested use of this reagent is 0.25 - 1.0 µg per ml (1:500 - 1:2000). It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

This clone may require prolonged exposure times to visualize proteins by western blot. We strongly recommend loading at least 30 µg of protein to facilitate detection.

Application References

(PubMed link indicates BioLegend citation)
  1. Moroishi T, et al. 2014. J Biol Chem. 289: 16430. (WB)
  2. Chollangi S, et al. 2012. J Biol Chem. 287: 23710. (WB)
RRID
AB_2749875 (BioLegend Cat. No. 672603)
AB_2749875 (BioLegend Cat. No. 672604)

Antigen Details

Structure
FBXL5 is a 691 amino acid protein with a predicted molecular weight of 78.6 kD. The protein contains an N-terminal hemerythrin like domain, followed by a F-box domain and seven leurcine-rich repeats.
Distribution

Cytosol and nucleus; ubiquitous expression

Function
Systemic and cellular iron homeostasis
Interaction
IRP2, IRP1, SKP1, Cullin-1, RBX1, HERC2
Ligand/Receptor
Iron and oxygen
Biology Area
Cell Biology, Ubiquitin/Protein Degradation
Antigen References
  1. Johnson NB, et al. 2017. J. Biol. Chem. 292: 15976
  2. Vinas-Castells R, et al. 2014. Nucleic Acids Res. 42: 1079
  3. Moroishi T, et al. 2014. J Biol Chem. 289: 16430.
  4. Ruiz JC, et al. 2013. J Biol Chem 288:552.
  5. Chollangi S, et al. 2012. J Biol Chem. 287: 23710.
  6. Moroishi T, et al. 2011. Cell Metab. 14:339
  7. Salahudeen AA, et al. 2009. Science. 326: 722
  8. Vashisht AA, et al. 2009. Science. 326: 718
Gene ID
2634 View all products for this Gene ID
UniProt
View information about FBXL5 on UniProt.org
Go To Top Version: 1    Revision Date: 07/25/2018

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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