Anti-Ferritin 抗体 [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 (Intra), WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-Ferritin antibody [EPR3004Y]
Ferritin 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR3004Y] to Ferritin -
由来種
Rabbit -
特異性
We recommend following our protocol when testing ICC and recommend using lower dilution in samples having low expression level of Ferritin.
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アプリケーション
適用あり: ICC/IF, Flow Cyt (Intra), WBmore details
適用なし: IHC-P -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide within Human Ferritin aa 50-150. The exact sequence is proprietary.
Database link: P02794 -
ポジティブ・コントロール
- WB: HEK293T, Jurkat, SH-SY-5Y, C6 and Raw264.7 cell lysates; Human and rat brain lysates ICC/IF: Jurkat cells Flow Cyt (intra): SH-SY5Y cells
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特記事項
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.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
バッファー
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR3004Y -
アイソタイプ
IgG -
研究分野
関連製品
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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
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab75973の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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ICC/IF |
1/100.
We recommend following our protocol when testing ICC and recommend using lower dilution in samples having low expression level of Ferritin. |
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Flow Cyt (Intra) |
1/50.
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WB | (6) |
1/1000 - 1/2000. Detects a band of approximately 19, 21 kDa (predicted molecular weight: 21,19 kDa).
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特記事項 |
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ICC/IF
1/100. We recommend following our protocol when testing ICC and recommend using lower dilution in samples having low expression level of Ferritin. |
Flow Cyt (Intra)
1/50. |
WB
1/1000 - 1/2000. Detects a band of approximately 19, 21 kDa (predicted molecular weight: 21,19 kDa). |
ターゲット情報
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機能
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. -
配列類似性
Belongs to the ferritin family.
Contains 1 ferritin-like diiron domain. - Information by UniProt
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参照データベース
- 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 -
製品の状態
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. -
別名
- Cell proliferation-inducing gene 15 protein antibody
- Ferritin H subunit antibody
- Ferritin heavy chain antibody
see all
画像
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All lanes : Anti-Ferritin antibody [EPR3004Y] (ab75973) at 1/1000 dilution
Lane 1 : Wild-type MCF7 cell lysate
Lane 2 : FTH1 knockout MCF7 cell lysate
Lane 3 : Wild-type HEK-293T ab255553 cell lysate
Lane 4 : FTH1 knockout HEK-293T ab260185 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 21,19 kDa
Observed band size: 24 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Ferritin antibody [EPR3004Y] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab75973 was shown to bind specifically to Ferritin. A band was observed at 24 kDa in wild-type MCF7 cell lysates with no signal observed at this size in FTH1 knockout cell line ab269493 (knockout cell lysate ab269655). To generate this image, wild-type and FTH1 knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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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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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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Intracellular 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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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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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 -
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?
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (86)
ab75973 は 86 報の論文で使用されています。
- Laferrière F et al. Similar neuronal imprint and no cross-seeded fibrils in α-synuclein aggregates from MSA and Parkinson's disease. NPJ Parkinsons Dis 8:10 (2022). PubMed: 35027576
- Rosato BE et al. SEC23B Loss-of-Function Suppresses Hepcidin Expression by Impairing Glycosylation Pathway in Human Hepatic Cells. Int J Mol Sci 23:N/A (2022). PubMed: 35163229
- Wu X et al. Altered expression of ferroptosis markers and iron metabolism reveals a potential role of ferroptosis in vitiligo. Pigment Cell Melanoma Res 35:328-341 (2022). PubMed: 35218147
- Soliman SSM et al. Novel Secreted Peptides From Rhizopus arrhizus var. delemar With Immunomodulatory Effects That Enhance Fungal Pathogenesis. Front Microbiol 13:863133 (2022). PubMed: 35387075
- Wang Y et al. α-Lipoic Acid-Plus Ameliorates Endothelial Injury by Inhibiting the Apoptosis Pathway Mediated by Intralysosomal Cathepsins in an In Vivo and In Vitro Endothelial Injury Model. Oxid Med Cell Longev 2022:8979904 (2022). PubMed: 35450412