Cat. # | Size | Price | Inventory |
---|---|---|---|
48322S | 100 µl |
REACTIVITY | H |
SENSITIVITY | Endogenous |
MW (kDa) | 62 |
Source/Isotype | Rabbit IgG |
Product Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Immunohistochemistry (Paraffin) | 1:125 - 1:500 |
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Loading of prestained molecular weight markers (#59329, 10 µl/lane) to verify electrotransfer and biotinylated protein ladder (#7727, 10 µl/lane) to determine molecular weights are recommended.
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: Do not allow slides to dry at any time during this procedure.
For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; follow with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min.
RECOMMENDED DETECTION REAGENTS |
SignalStain® Boost IHC Detection Reagent (HRP, Rabbit) #8114 | SignalStain® Boost IHC Detection Reagent (AP, Rabbit) #18653 |
---|---|---|
COMPATIBLE CHROMOGEN |
SignalStain® DAB Substrate Kit #8059 | SignalStain® Vibrant Red Alkaline Phosphatase Substrate Kit #76713 |
SignalStain® Vivid Purple Peroxidase Substrate Kit #96632 | SignalStain® Ultra Blue Alkaline Phosphatase Substrate Kit #12824 | |
SignalStain® Deep Black Peroxidase Substrate Kit #72986 | ||
SignalStain® Radiant Yellow Peroxidase Substrate Kit #69644 |
NOTE: Use of detection reagents other than those specified in this protocol may require further optimization of the primary antibody to account for the different sensitivities of the detection reagents.
posted February 2010
revised June 2020
Protocol Id: 283
Human
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu265 of human SLC40A1/Ferroportin-1 protein.
Ferroportin-1 (FPN1), also known as solute carrier family 40 member 1 (SLC40A1) or iron-regulated transporter 1 (IREG1), is a multi-pass membrane protein that transports iron from the inside of the cell into the blood. As the only known iron exporter, FPN1 is critical for maintaining systemic iron homeostasis (1). FPN1 activity is regulated by hepcidin, a peptide hormone secreted by the liver in response to iron loading and inflammation. Circulating hepcidin binds to FPN1 and induces its internalization and degradation, thereby reducing iron efflux to the plasma (2,3). Expression of FPN1 may also be regulated post-transcriptionally by several microRNAs that directly target its 3' untranslated region (4-6). Mutations in the SLC40A1 gene encoding FPN1 are associated with type IV hemochromatosis (7) as well as several neural tube and patterning defects, including spina bifida, exencephaly, and forebrain truncations (8). Loss of FPN1 in the brains of Alzheimer's disease (AD) mouse models and patients has been shown to promote ferroptosis, leading to neuronal death and memory impairment. Therefore, targeting of FPN1 may be a promising therapeutic approach for AD (9).
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