Recombinant Anti-Ferritin antibody [EPR3004Y] (ab75973)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3004Y] to Ferritin
- Suitable for: ICC/IF, Flow Cyt, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-Ferritin antibody [EPR3004Y]
See all Ferritin primary antibodies -
Description
Rabbit monoclonal [EPR3004Y] to Ferritin -
Host species
Rabbit -
Specificity
In human, this antibody recognizes both, Ferritin heavy and light chains. In mouse and rat, it only recognizes Ferritin heavy chain.
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Tested applications
Suitable for: ICC/IF, Flow Cyt, WBmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human Ferritin aa 50-150. The exact sequence is proprietary.
Database link: P02794 -
Positive control
- WB: HEK293T, Jurkat, SH-SY-5Y, C6 and Raw264.7 cell lysates; Human and rat brain lysates ICC/IF: Jurkat cells Flow Cyt: SH-SY5Y cells
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3004Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
Applications
Our Abpromise guarantee covers the use of ab75973 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| ICC/IF | 1/100. | |
| Flow Cyt | 1/50. | |
| WB | 1/1000 - 1/2000. Detects a band of approximately 19, 21 kDa (predicted molecular weight: 21,19 kDa). |
Target
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Function
Stores iron in a soluble, non-toxic, readily available form. Important for iron homeostasis. Has ferroxidase activity. Iron is taken up in the ferrous form and deposited as ferric hydroxides after oxidation. Also plays a role in delivery of iron to cells. Mediates iron uptake in capsule cells of the developing kidney. -
Sequence similarities
Belongs to the ferritin family.
Contains 1 ferritin-like diiron domain. - Information by UniProt
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Database links
- Entrez Gene: 2495 Human
- Entrez Gene: 2512 Human
- Entrez Gene: 14319 Mouse
- Entrez Gene: 14325 Mouse
- Entrez Gene: 25319 Rat
- Entrez Gene: 29292 Rat
- Omim: 608847 Human
- SwissProt: P02792 Human
see all -
Form
Subunit structure: Oligomer of 24 subunits. There are two types of subunits: L (light) chain and H (heavy) chain. The major chain can be light or heavy, depending on the species and tissue type. In the human liver, the heavy chain is predominant. The functional molecule forms a roughly spherical shell with a diameter of 12 nm and contains a central cavity into which the insoluble mineral iron core is deposited. -
Alternative names
- Cell proliferation-inducing gene 15 protein antibody
- Ferritin H subunit antibody
- Ferritin heavy chain antibody
see all
Images
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![Western blot - Anti-Ferritin antibody [EPR3004Y] (ab75973)](https://www.abcam.com/ps/products/75/ab75973/Images/ab75973-393623-anti-ferritin-antibody-epr3004y-western-blot-human-knockout-lysate.jpg)
Western blot - Anti-Ferritin antibody [EPR3004Y] (ab75973)All lanes : Anti-Ferritin antibody [EPR3004Y] (ab75973) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : FTH1 knockout HEK293T cell lysate
Lane 3 : SH-SY5Y cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 21,19 kDa
Observed band size: 21 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab75973 observed at 21 kDa. Red - loading control ab8245 observed at 36 kDa.
ab75973 Anti-Ferritin antibody [EPR3004Y] was shown to specifically react with Ferritin in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266581 (knockout cell lysate ab256924) was used. Wild-type and Ferritin knockout samples were subjected to SDS-PAGE. ab75973 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Immunocytochemistry/ Immunofluorescence - Anti-Ferritin antibody [EPR3004Y] (ab75973)](https://www.abcam.com/ps/products/75/ab75973/Images/ab75973-2-ab75973ICCIF1.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Ferritin antibody [EPR3004Y] (ab75973)Immunofluorescence staining of Jurkat cells with purified ab75973 at a working dilution of 1/100, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 100% methanol and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab75973 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
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![Flow Cytometry - Anti-Ferritin antibody [EPR3004Y] (ab75973)](https://www.abcam.com/ps/products/75/ab75973/Images/ab75973-8-anti-ferritin-antibody-epr3004y-flow-cytometry-sh-sy5y1.jpg)
Flow Cytometry - Anti-Ferritin antibody [EPR3004Y] (ab75973)Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized SH-SY5Y (Human neuroblastoma epithelial cell) cells labelling Ferritin with ab75973 at 1/50 dilution (1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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![Western blot - Anti-Ferritin antibody [EPR3004Y] (ab75973)](https://www.abcam.com/ps/products/75/ab75973/Images/ab75973-395640-AntiFerritin-antibody.jpg)
Western blot - Anti-Ferritin antibody [EPR3004Y] (ab75973)All lanes : Anti-Ferritin antibody [EPR3004Y] (ab75973) at 1/2000 dilution
Lane 1 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates
Lane 2 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates
Lane 3 : C6 (Rat glial tumor glial cell) whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/20000 dilution
Predicted band size: 21,19 kDaObserved MW: 21 kDA
Blocking/diluting buffer and concentration: 5% NFDM/TBST
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![Western blot - Anti-Ferritin antibody [EPR3004Y] (ab75973)](https://www.abcam.com/ps/products/75/ab75973/Images/ab75973-256964-ab75973WB3.jpg)
Western blot - Anti-Ferritin antibody [EPR3004Y] (ab75973)Anti-Ferritin antibody [EPR3004Y] (ab75973) at 1/10000 dilution (purified) + Raw264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 21,19 kDa
Observed band size: 21 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
![Western blot - Anti-Ferritin antibody [EPR3004Y] (ab75973)](https://www.abcam.com/ps/products/75/ab75973/Images/ab75973.jpg)
Western blot - Anti-Ferritin antibody [EPR3004Y] (ab75973)All lanes : Anti-Ferritin antibody [EPR3004Y] (ab75973) at 1/2000 dilution (Unpurified)
Lane 1 : Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate
Lane 2 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) cell lysate
Lane 3 : Human brain lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 21,19 kDa
Observed band size: 19,21 kDa why is the actual band size different from the predicted?
Protocols
References (36)
ab75973 has been referenced in 36 publications.
- Zhao H et al. Long-term voluntary running prevents the onset of symptomatic Friedreich's ataxia in mice. Sci Rep 10:6095 (2020). WB ; Mouse . PubMed: 32269244
- Nagpal A et al. Neoadjuvant neratinib promotes ferroptosis and inhibits brain metastasis in a novel syngeneic model of spontaneous HER2+ve breast cancer metastasis. Breast Cancer Res 21:94 (2019). PubMed: 31409375
- Okuno K et al. Haploinsufficiency of Transferrin Receptor 1 Impairs Angiogenesis with Reduced Mitochondrial Complex I in Mice with Limb Ischemia. Sci Rep 9:13658 (2019). PubMed: 31541184
- Zhang L et al. Role of mitochondrial calcium uniporter-mediated Ca2+ and iron accumulation in traumatic brain injury. J Cell Mol Med 23:2995-3009 (2019). PubMed: 30756474
- Yu X et al. Iron Transport from Ferrous Bisglycinate and Ferrous Sulfate in DMT1-Knockout Human Intestinal Caco-2 Cells. Nutrients 11:N/A (2019). PubMed: 30813537
