Anti-FOXO3A 抗体 (ab23683)
Key features and details
- Rabbit polyclonal to FOXO3A
- Suitable for: WB, IHC-P, ICC/IF
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
リコンビナント抗体で、ロット間での高い再現性を実現
- 異なるロット間での安定した再現性
- 容易なスケールアップ
- 評価試験による特異性の確認済み
- 倫理基準に準拠 - アニマル・フリーの生産
製品の概要
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製品名
Anti-FOXO3A antibody
FOXO3A 一次抗体 製品一覧 -
製品の詳細
Rabbit polyclonal to FOXO3A -
由来種
Rabbit -
特異性
Replenishment batches of our polyclonal antibody, ab23683 are tested in WB. Previous batches were additionally validated in ICC/IF and IHC-P. These applications are still expected to work and are covered by our Abpromise guarantee. You may also be interested in our alternative recombinant antibody, ab109629.
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アプリケーション
適用あり: WB, IHC-P, ICC/IFmore details -
種交差性
交差種: Mouse, Rat, Human
交差が予測される動物種: Dog -
免疫原
Synthetic peptide corresponding to Human FOXO3A aa 650 to the C-terminus conjugated to keyhole limpet haemocyanin.
(Peptide available asab24031) -
ポジティブ・コントロール
- Recombinant Human FOXO3A protein (ab114191) can be used as a positive control in WB. This antibody gave a positive signal in the following whole cell lysates: Jurkat (Human T cell lymphoblast-like cell line) This antibody gave a positive signal in the following tissue lysates: Mouse Heart Mouse Skeletal Muscle Rat Heart This antibody gave a positive result in IHC in the following FFPE tissue: Human normal lung.
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特記事項
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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 or -80°C. Avoid freeze / thaw cycle. -
バッファー
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading...
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精製度
Immunogen affinity purified -
ポリ/モノ
ポリクローナル -
アイソタイプ
IgG -
研究分野
関連製品
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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
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Positive Controls
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Recombinant Protein
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ab23683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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WB |
Use a concentration of 1 µg/ml. Detects a band of approximately 90 kDa (predicted molecular weight: 71 kDa).
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IHC-P |
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF |
Use a concentration of 5 µg/ml.
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特記事項 |
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WB
Use a concentration of 1 µg/ml. Detects a band of approximately 90 kDa (predicted molecular weight: 71 kDa). |
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
Use a concentration of 5 µg/ml. |
ターゲット情報
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機能
Transcriptional activator which triggers apoptosis in the absence of survival factors, including neuronal cell death upon oxidative stress. Recognizes and binds to the DNA sequence 5'-[AG]TAAA[TC]A-3'. -
組織特異性
Ubiquitous. -
関連疾患
Note=A chromosomal aberration involving FOXO3 is found in secondary acute leukemias. Translocation t(6;11)(q21;q23) with MLL/HRX. -
配列類似性
Contains 1 fork-head DNA-binding domain. -
翻訳後修飾
In the presence of survival factors such as IGF-1, phosphorylated on Thr-32 and Ser-253 by AKT1/PKB. This phosphorylated form then interacts with 14-3-3 proteins and is retained in the cytoplasm. Survival factor withdrawal induces dephosphorylation and promotes translocation to the nucleus where the dephosphorylated protein induces transcription of target genes and triggers apoptosis. Although AKT1/PKB doesn't appear to phosphorylate Ser-315 directly, it may activate other kinases that trigger phosphorylation at this residue. Phosphorylated by STK4 on Ser-209 upon oxidative stress, which leads to dissociation from YWHAB/14-3-3-beta and nuclear translocation. Phosphorylated by PIM1. -
細胞内局在
Cytoplasm > cytosol. Nucleus. Translocates to the nucleus upon oxidative stress and in the absence of survival factors. - Information by UniProt
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参照データベース
- Entrez Gene: 481954 Dog
- Entrez Gene: 2309 Human
- Entrez Gene: 56484 Mouse
- Entrez Gene: 294515 Rat
- Omim: 602681 Human
- SwissProt: O43524 Human
- SwissProt: Q9WVH4 Mouse
- Unigene: 220950 Human
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別名
- AF6q21 antibody
- AF6q21 protein antibody
- DKFZp781A0677 antibody
see all
画像
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All lanes : Anti-FOXO3A antibody (ab23683) at 1 µg/ml
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 :Human FOXO3 (FOXO3A) knockout HEK-293T cell lysate (ab256922)
Lane 3 : Wild-type HEK-293 cell lysate
Lane 4 :Human FOXO3 (FOXO3A) knockout HEK-293 cell lysate (ab261649)
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 71 kDa
Observed band size: 82 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab23683 observed at 82 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab23683 was shown to react with FOXO3A in wild-type cells in Western blot with loss of signal observed in FOXO3 knockout cell lines. Wild-type and FOXO3 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab23683 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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ICC/IF image of ab23683 stained MCF7 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab23683, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% HepG2 fixed (10 min) HepG2 cells at 5µg/ml.
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IHC image of FOXO3A staining in Human normal lung formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab23683, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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All lanes : Anti-FOXO3A antibody (ab23683) at 1 µg/ml
Lane 1 : Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : FOXO3 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 71 kDa
Observed band size: 82 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab23683 observed at 71 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab23683 was shown to recognize FOXO3A in wild-type HEK-293 cells as signal was lost at the expected MW in FOXO3 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and FOXO3 knockout samples were subjected to SDS-PAGE. Ab23683 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-FOXO3A antibody (ab23683) at 1 µg/ml
Lane 1 :Jurkat whole cell lysate (ab7899)
Lane 2 :Jurkat whole cell lysate (ab7899) with Human FOXO3A peptide (ab24031) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Predicted band size: 71 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?
Additional bands at: 50 kDa (possible cleavage fragment), 50 kDa (possible cross reactivity)
ab23683 detects a band at approximately 90 kDa that corresponds in size to that seen for FOXO3A. This protein has been shown to migrate at a size larger than the predicted molecular weight (see Yin et al., J. Biol. Chem., Vol. 279, Issue 44, 45721-45727). It also detects a band at 50 kDa which could be a cleavage fragment. Both bands are partially blocked by the addition of the immunizing peptide. -
All lanes : Anti-FOXO3A antibody (ab23683) at 1 µg/ml
Lane 1 : Heart (Mouse) Tissue Lysate - normal tissue
Lane 2 :Mouse skeletal muscle tissue lysate - total protein (ab29711)
Lane 3 : Heart (Rat) Tissue Lysate - normal tissue
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Predicted band size: 71 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?
Additional bands at: 16 kDa, 27 kDa. We are unsure as to the identity of these extra bands.ab23683 detects a band at approximately 90 kDa that corresponds in size to that seen for FOXO3A. This protein has been shown to migrate at a size larger than the predicted molecular weight (see Yin et al., J. Biol. Chem., Vol. 279, Issue 44, 45721-45727).
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 (24)
ab23683 は 24 報の論文で使用されています。
- Meng L et al. MeCP2 inhibits ischemic neuronal injury by enhancing methylation of the FOXO3a promoter to repress the SPRY2-ZEB1 axis. Exp Mol Med 54:1076-1085 (2022). PubMed: 35915222
- Sezginer P et al. The effect of controlled ovarian hyperstimulation on ovarian reserve via PTEN pathway. Reprod Fertil 3:187-97 (2022). PubMed: 35972314
- Cieminski K et al. Swim training affects Akt signaling and ameliorates loss of skeletal muscle mass in a mouse model of amyotrophic lateral sclerosis. Sci Rep 11:20899 (2021). PubMed: 34686697
- Sun H et al. Cardamonin Promotes the Apoptosis and Chemotherapy Sensitivity to Gemcitabine of Pancreatic Cancer Through Modulating the FOXO3a-FOXM1 Axis. Dose Response 19:15593258211042163 (2021). PubMed: 34987330
- Ai Y et al. Exosomal LncRNA LBX1-AS1 Derived From RBPJ Overexpressed-Macrophages Inhibits Oral Squamous Cell Carcinoma Progress via miR-182-5p/FOXO3. Front Oncol 11:605884 (2021). PubMed: 33816238