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Product Info Summary
| SKU: | A00506-5 |
|---|---|
| Size: | 100 µg/vial |
| Reactive Species: | Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-Hfe Antibody Picoband®
SKU/Catalog Number
A00506-5
Size
100 µg/vial
Form
Lyophilized
Description
Boster Bio Anti-Hfe Antibody Picoband® catalog # A00506-5. Tested in ELISA, Flow Cytometry, IHC, WB applications. This antibody reacts with Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.
Cite This Product
Anti-Hfe Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00506-5)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived mouse Hfe recombinant protein (Position: N112-H227).
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00506-5 is reactive to Hfe in Mouse, Rat
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.
Observed Molecular Weight
40-58 kDa
Calculated molecular weight
40108 MW
Background of HFE
Human homeostatic iron regulator protein, also known as the HFE protein (High FE2+), is a protein which in humans is encoded by the HFE gene. Enables transferrin receptor binding activity. Involved in positive regulation of gene expression. Acts upstream of or within hormone biosynthetic process and multicellular organismal iron ion homeostasis. Part of HFE-transferrin receptor complex. Is expressed in brain; choroid invagination; diencephalon roof plate; medulla oblongata part of 4th ventricle choroid plexus; and metencephalon part of 4th ventricle choroid plexus. Used to study hemochromatosis type 1. Human ortholog(s) of this gene implicated in several diseases, including acute porphyria (multiple); arthritis (multiple); bone marrow cancer (multiple); hemochromatosis (multiple); and liver disease (multiple). Orthologous to human HFE (homeostatic iron regulator).
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A00506-5 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.25-0.5 μg/ml, Mouse, Rat
Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Rat
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Mouse
ELISA, 0.1-0.5 μg/ml, -
Positive Control
WB: rat liver tissue, mouse brain tissue, mouse liver tissue, mouse Neuro-2a whole cell
IHC: rat liver tissue
FCM: HEPA1-6 cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of Hfe using anti-Hfe antibody (A00506-5).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: rat liver tissue lysates,
Lane 2: mouse brain tissue lysates,
Lane 3: mouse liver tissue lysates,
Lane 4: mouse Neuro-2a whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Hfe antigen affinity purified polyclonal antibody (Catalog # A00506-5) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Hfe at approximately 40-58 kDa. The expected band size for Hfe is at 41 kDa.
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Figure 2. IHC analysis of Hfe using anti-Hfe antibody (A00506-5).
Hfe was detected in a paraffin-embedded section of rat liver tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Hfe Antibody (A00506-5) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
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Figure 3. Flow Cytometry analysis of HEPA1-6 cells using anti-Hfe antibody (A00506-5).
Overlay histogram showing HEPA1-6 cells stained with A00506-5 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-Hfe Antibody (A00506-5, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Protein Target Info & Infographic
Gene/Protein Information For Hfe (Source: Uniprot.org, NCBI)
Gene Name
Hfe
Full Name
Hereditary hemochromatosis protein
Weight
40108 MW
Superfamily
MHC class I family
Alternative Names
Hereditary hemochromatosis protein;HLA-H;HFE;HLAH; HFE HFE1, HH, HLA-H, MVCD7, TFQTL2 homeostatic iron regulator hereditary hemochromatosis protein|MHC class I-like protein HFE|hereditary hemochromatosis protein HLA-H|high Fe
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on Hfe, check out the Hfe Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for Hfe: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-Hfe Antibody Picoband® (A00506-5)
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