Anti-DIAPH1 抗体 [EPR7948] - BSA and Azide free (ab248325)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR7948] to DIAPH1 - BSA and Azide free
- Suitable for: IHC-P, WB, ICC/IF, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free
DIAPH1 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR7948] to DIAPH1 - BSA and Azide free -
由来種
Rabbit -
アプリケーション
適用あり: IHC-P, WB, ICC/IF, Flow Cyt (Intra)more details -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- WB: HCT116 and 293T cell lysates' HEK-293, RAW 264.7 and Human brain lysates, PC-12 and MCF7 lysates; Flow Cyt (intra): HeLa cells; ICC/IF: HeLa cells; IHC-P: Human, rat, and mouse kidney tissue sections.
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特記事項
ab248325 is the carrier-free version of ab129167.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. Do Not Freeze. -
バッファー
pH: 7.2
Constituent: PBS -
キャリア・フリー
はい -
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
EPR7948 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
- Anti-DIAPH1 antibody [EPR7948] (ab129167)
- Alexa Fluor® 647 Anti-DIAPH1 antibody [EPR7948] (ab207061)
- PE Anti-DIAPH1 antibody [EPR7948] (ab306191)
- APC Anti-DIAPH1 antibody [EPR7948] (ab306192)
- HRP Anti-DIAPH1 antibody [EPR7948] (ab306193)
- Alexa Fluor® 488 Anti-DIAPH1 antibody [EPR7948] (ab309647)
- Alexa Fluor® 594 Anti-DIAPH1 antibody [EPR7948] (ab310380)
- Alexa Fluor® 555 Anti-DIAPH1 antibody [EPR7948] (ab311905)
- Alexa Fluor® 568 Anti-DIAPH1 antibody [EPR7948] (ab312376)
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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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Positive Controls
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ab248325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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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 150 kDa (predicted molecular weight: 141 kDa).
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ICC/IF |
Use at an assay dependent concentration.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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特記事項 |
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IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
WB
Use at an assay dependent concentration. Detects a band of approximately 150 kDa (predicted molecular weight: 141 kDa). |
ICC/IF
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
ターゲット情報
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機能
Acts in a Rho-dependent manner to recruit PFY1 to the membrane. Required for the assembly of F-actin structures, such as actin cables and stress fibers. Nucleates actin filaments. Binds to the barbed end of the actin filament and slows down actin polymerization and depolymerization. Required for cytokinesis, and transcriptional activation of the serum response factor. DFR proteins couple Rho and Src tyrosine kinase during signaling and the regulation of actin dynamics. Functions as a scaffold protein for MAPRE1 and APC to stabilize microtubules and promote cell migration (By similarity). Has neurite outgrowth promoting activity (By similarity). In hear cells, it may play a role in the regulation of actin polymerization in hair cells. The MEMO1-RHOA-DIAPH1 signaling pathway plays an important role in ERBB2-dependent stabilization of microtubules at the cell cortex. It controls the localization of APC and CLASP2 to the cell membrane, via the regulation of GSK3B activity. In turn, membrane-bound APC allows the localization of the MACF1 to the cell membrane, which is required for microtubule capture and stabilization. -
組織特異性
Expressed in brain, heart, placenta, lung, kidney, pancreas, liver, skeletal muscle and cochlea. -
関連疾患
Defects in DIAPH1 are the cause of deafness autosomal dominant type 1 (DFNA1) [MIM:124900]. DFNA1 is a form of sensorineural hearing loss. Sensorineural deafness results from damage to the neural receptors of the inner ear, the nerve pathways to the brain, or the area of the brain that receives sound information. -
配列類似性
Belongs to the formin homology family. Diaphanous subfamily.
Contains 1 DAD (diaphanous autoregulatory) domain.
Contains 1 FH1 (formin homology 1) domain.
Contains 1 FH2 (formin homology 2) domain.
Contains 1 GBD/FH3 (Rho GTPase-binding/formin homology 3) domain. -
ドメイン
DRFs are regulated by intramolecular GBD-DAD binding where Rho-GTP activates the DRFs by disrupting the GBD-DAD interaction (By similarity). DCAF7 binds to the FH2 (formin homology 2) domain. -
細胞内局在
Cell membrane. Cell projection > ruffle membrane. Cytoplasm > cytoskeleton. Membrane ruffles, especially at the tip of ruffles, of motile cells. - Information by UniProt
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参照データベース
- Entrez Gene: 1729 Human
- Entrez Gene: 13367 Mouse
- Entrez Gene: 307483 Rat
- Omim: 602121 Human
- SwissProt: O60610 Human
- SwissProt: O08808 Mouse
- Unigene: 529451 Human
- Unigene: 195916 Mouse
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別名
- DIAPH1 antibody
- deafness, autosomal dominant 1 antibody
- DFNA1 antibody
see all
画像
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All lanes : Anti-DIAPH1 antibody [EPR7948] (ab129167) at 1/10000 dilution (Purified)
Lane 1 : PC-12 (Rat adrenal gland pheochromocytoma ) whole cell lysate
Lane 2 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 141 kDa -
All lanes : Anti-DIAPH1 antibody [EPR7948] (ab129167) at 1/1000 dilution (Purified)
Lane 1 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate
Lane 2 : Human brain lysate
Lane 3 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 141 kDa -
All lanes : Anti-DIAPH1 antibody [EPR7948] (ab129167) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : DIAPH1 knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 141 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab129167).
Lanes 1- 2: Merged signal (red and green). Green - ab129167 observed at 150 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab129167 was shown to react with DIAPH1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266120 (knockout cell lysate ab257411) was used. Wild-type HEK-293T and DIAPH1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab129167 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 1000 dilution and a 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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This data was developed using ab129167, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling DIAPH1 with Purified ab129167 at 1/50 dilution (10 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 dilution (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/mL). DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control. -
This data was developed using ab129167, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue sections labeling DIAPH1 with Purified ab129167 at 1/100 dilution (5.96 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain. -
This data was developed using ab129167, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling DIAPH1 with Purified ab129167 at 1/100 dilution (5.96 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain. -
This data was developed using ab129167, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue sections labeling DIAPH1 with Purified ab129167 at 1/100 dilution (5.96 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain. -
This data was developed using ab129167, the same antibody clone in a different buffer formulation.
Intracellular Flow Cytometry analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling DIAPH1 with purified ab129167 at 1/50 dilution. Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 was used as the secondary antibody (red). Rabbit monoclonal IgG (ab172730) was used as the isotype control (black). Cells without incubation with primary and secondary antibodies were used as the unlabeled control (blue).
プロトコール
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
データシートおよび資料
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Datasheet download
Certificate of Compliance
参考文献 (0)
ab248325 は論文での使用が確認できていません。