Anti-AMF 抗体 [1B7D7] (ab66340)
![Western blot - Anti-AMF antibody [1B7D7] (ab66340)](https://www.abcam.co.jp/ps/products/66/ab66340/Images/ab66340-454787-anti-amf-antibody-1b7d7-western-blot-wildtype-hek293t-gpi-knockout-hek293t.jpg "Western blot - Anti-AMF antibody [1B7D7] (ab66340)")
Key features and details
- Mouse monoclonal [1B7D7] to AMF
- Suitable for: ICC/IF, IHC-P, WB, Flow Cyt
- Knockout validated
- Reacts with: Human
- Isotype: IgG1
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-AMF antibody [1B7D7]
AMF 一次抗体 製品一覧 -
製品の詳細
Mouse monoclonal [1B7D7] to AMF -
由来種
Mouse -
アプリケーション
適用あり: ICC/IF, IHC-P, WB, Flow Cytmore details -
種交差性
交差種: Human -
免疫原
Recombinant fragment corresponding to Human AMF.
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ポジティブ･コントロール
- WB: HEK293T, HepG2 and SMMC-7721 cell lysate. Flow Cyt: HepG2 cells. ICC: L-02 cells. IHC-P: Human cerebral cortex tissue.
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特記事項
This product was changed from ascites to supernatant. Lot no’s high than GR185888-22 are from Tissue Culture Supernatant.
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. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
バッファー
Preservative: 0.05% Sodium azide
Constituent: PBS -
Concentration information loading... -
精製度
Protein G purified -
特記事項（精製）
Purified from TCS. -
ポリ/モノ
モノクローナル -
クローン名
1B7D7 -
アイソタイプ
IgG1 -
研究分野
関連製品
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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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ab66340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 |
|---|---|---|
| ICC/IF | (1) |
1/200 - 1/1000.
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| IHC-P | (1) |
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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| WB | (2) |
1/500 - 1/5000. Predicted molecular weight: 63 kDa.
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| Flow Cyt |
1/100.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
| 特記事項 |
|---|
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ICC/IF
1/200 - 1/1000. |
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IHC-P
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
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WB
1/500 - 1/5000. Predicted molecular weight: 63 kDa. |
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Flow Cyt
1/100. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
ターゲット情報
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機能
Besides it's role as a glycolytic enzyme, mammalian GPI can function as a tumor-secreted cytokine and an angiogenic factor (AMF) that stimulates endothelial cell motility. GPI is also a neurotrophic factor (Neuroleukin) for spinal and sensory neurons. -
パスウェイ
Carbohydrate degradation; glycolysis; D-glyceraldehyde 3-phosphate and glycerone phosphate from D-glucose: step 2/4. -
関連疾患
Defects in GPI are the cause of hemolytic anemia non-spherocytic due to glucose phosphate isomerase deficiency (HA-GPID) [MIM:613470]. It is a form of anemia in which there is no abnormal hemoglobin or spherocytosis. It is caused by glucose phosphate isomerase deficiency. Severe GPI deficiency can be associated with hydrops fetalis, immediate neonatal death and neurological impairment. -
配列類似性
Belongs to the GPI family. -
翻訳後修飾
Phosphorylation at Ser-185 by CK2 has been shown to decrease enzymatic activity and may contribute to secretion by a non-classical secretory pathway.
ISGylated. -
細胞内局在
Cytoplasm. Secreted. - Information by UniProt
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参照データベース
- Entrez Gene: 2821 Human
- Omim: 172400 Human
- SwissProt: P06744 Human
- Unigene: 466471 Human
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別名
- AMF antibody
- Aurocrine motility factor antibody
- Autocrine motility factor antibody
see all
画像
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Western blot - Anti-AMF antibody [1B7D7] (ab66340)All lanes : Anti-AMF antibody [1B7D7] (ab66340) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : GPI knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 63 kDa
Observed band size: 63 kDaLanes 1 - 2: Merged signal (red and green). Green - ab66340 observed at 63 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37 kDa.
ab66340 was shown to react with AMF in wild-type HEK-293T cells in Western blot with loss of signal observed in GPI knockout cell line ab266834 (GPI knockout cell lysate ab257458). Wild-type HEK-293T and GPI knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab66340 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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![Flow Cytometry - Anti-AMF antibody [1B7D7] (ab66340)](https://www.abcam.co.jp/ps/products/66/ab66340/Images/ab66340-195306-anti-glucose-6-phosphate-isomerase-antibody-1b7d7-flow-cytometry.jpg)
Flow Cytometry - Anti-AMF antibody [1B7D7] (ab66340)Overlay histogram showing HepG2 cells stained with ab66340 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab66340, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H+L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HepG2 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions. -
![Immunocytochemistry/ Immunofluorescence - Anti-AMF antibody [1B7D7] (ab66340)](https://www.abcam.co.jp/ps/products/66/ab66340/Images/ab66340-80124-anti-glucose-6-phosphate-isomerase-antibody-1b7d7-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-AMF antibody [1B7D7] (ab66340)ab66340 at 1000 dilution staining AMF in L-02 cells by Immunocytochemistry/ Immunofluorescence. An Alexa Fluor® 488 conjugated Goat polyclonal to mouse IgG1 was used as secondary antibody. Green staining in image show positive staining with ab66340, actin filaments were stained red with DY-554 phalloidin and nuclei stained blue with DRAQ5 fluorescent DNA dye.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMF antibody [1B7D7] (ab66340)](https://www.abcam.co.jp/ps/products/66/ab66340/Images/ab66340-80123-anti-glucose-6-phosphate-isomerase-antibody-1b7d7-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMF antibody [1B7D7] (ab66340)ab66340 (1µg/ml) staining AMF in human cerebral cortex using an automated system (DAKO Autostainer Plus). Using this protocol there is strong cytoplasmic staining of neurons and of the neuropil.
Inset panel depicts negative control (no primary antibody).
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required. -
![Western blot - Anti-AMF antibody [1B7D7] (ab66340)](https://www.abcam.co.jp/ps/products/66/ab66340/Images/ab66340-80125-anti-glucose-6-phosphate-isomerase-antibody-1b7d7-western-blot.jpg)
Western blot - Anti-AMF antibody [1B7D7] (ab66340)All lanes : Anti-AMF antibody [1B7D7] (ab66340) at 1/2000 dilution
Lane 1 : Cell lysates prepared from HepG2 cells.
Lane 2 : Cell lysates prepared from SMMC-7721 cells.
Lysates/proteins at 100 µg per lane.
Secondary
All lanes : HRP-conjugated Goat polyclonal to mouse IgG1
Predicted band size: 63 kDa
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 (19)
ab66340 は 19 報の論文で使用されています。
- Yang F et al. Circ-CTNNB1 drives aerobic glycolysis and osteosarcoma progression via m6A modification through interacting with RBM15. Cell Prolif 56:e13344 (2023). PubMed: 36181462
- Wei J et al. Osteoblasts induce glucose-derived ATP perturbations in chondrocytes through noncontact communication. Acta Biochim Biophys Sin (Shanghai) 54:625-636 (2022). PubMed: 35593470
- Zhao H et al. LINC01816 promotes the migration, invasion and epithelial-mesenchymal transition of thyroid carcinoma cells by sponging miR-34c-5p and regulating CRABP2 expression levels. Oncol Rep 45:N/A (2021). PubMed: 33786631
- Zhang Q et al. Supermeres are functional extracellular nanoparticles replete with disease biomarkers and therapeutic targets. Nat Cell Biol 23:1240-1254 (2021). PubMed: 34887515
- Weng ML et al. Fasting inhibits aerobic glycolysis and proliferation in colorectal cancer via the Fdft1-mediated AKT/mTOR/HIF1a pathway suppression. Nat Commun 11:1869 (2020). PubMed: 32313017