Anti-PBK/SPK 抗体 [EPR21983] (ab236872)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR21983] to PBK/SPK
- Suitable for: Flow Cyt (Intra), WB, IHC-P, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-PBK/SPK antibody [EPR21983]
PBK/SPK 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR21983] to PBK/SPK -
由来種
Rabbit -
アプリケーション
適用あり: Flow Cyt (Intra), WB, IHC-P, IPmore details -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Recombinant fragment. 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
- IHC-P: Human, rat and mouse testis tissue. Human gastric cancer tissue. WB: HeLa, HepG2, SW480, MCF7, A431, U-87 MG, Hepa1-6, C6, PC-12 and RAW 264.7 whole cell lysate. Human and rat testis lysate. Wild-type HAP1 cell lysate. IP: PBK/SPK IP in HepG2 whole cell lysate. Flow Cyt (intra): HeLa and HepG2 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 long term. Avoid freeze / thaw cycle. -
バッファー
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% 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
EPR21983 -
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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ChIP Related Products
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Compatible Secondaries
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Immunohistochemistry reagents
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Isotype control
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Recombinant Protein
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Related Products
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ab236872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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Flow Cyt (Intra) |
1/500.
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WB |
1/1000. Detects a band of approximately 38 kDa (predicted molecular weight: 36 kDa).
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IHC-P |
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IP |
1/30.
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特記事項 |
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Flow Cyt (Intra)
1/500. |
WB
1/1000. Detects a band of approximately 38 kDa (predicted molecular weight: 36 kDa). |
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IP
1/30. |
ターゲット情報
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機能
Phosphorylates MAP kinase p38. Seems to be active only in mitosis. May also play a role in the activation of lymphoid cells. When phosphorylated, forms a complex with TP53, leading to TP53 destabilization and attenuation of G2/M checkpoint during doxorubicin-induced DNA damage. -
組織特異性
Expressed in the testis and placenta. In the testis, restrictedly expressed in outer cell layer of seminiferous tubules. -
配列類似性
Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. MAP kinase kinase subfamily.
Contains 1 protein kinase domain. -
翻訳後修飾
Phosphorylated; in a cell-cycle dependent manner at mitosis. - Information by UniProt
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参照データベース
- Entrez Gene: 55872 Human
- Entrez Gene: 52033 Mouse
- Entrez Gene: 290326 Rat
- Omim: 611210 Human
- SwissProt: Q96KB5 Human
- SwissProt: Q9JJ78 Mouse
- Unigene: 104741 Human
- Unigene: 24337 Mouse
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別名
- Cancer/testis antigen 84 antibody
- CT84 antibody
- Epididymis luminal protein 164 antibody
see all
画像
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All lanes : Anti-PBK/SPK antibody [EPR21983] (ab236872) at 1/1000 dilution
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : PBK/SPK knockout HAP1 cell lysate
Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lane 4 : HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 36 kDa
Observed band size: 38 kDa why is the actual band size different from the predicted?
Exposure time: 15 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID:23547718; PMID:25909225).
ab236872 was shown to specifically react with PBK/SPK in wild-type HAP1 cells as signal was lost in PBK/SPK knockout cells. Wild-type and PBK/SPK knockout samples were subjected to SDS-PAGE. ab236872 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemical analysis of paraffin-embedded human testis tissue labeling PBK/SPK with ab236872 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in germ cells of human testis (PMID:16982762; PMID:25909225) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). -
All lanes : Anti-PBK/SPK antibody [EPR21983] (ab236872) at 1/1000 dilution
Lane 1 : SW480 (human colorectal adenocarcinoma epithelial cell), whole cell lysate
Lane 2 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate
Lane 3 : A431 (human epidermoid carcinoma epithelial cell), whole cell lysate
Lane 4 : U-87 MG (human glioblastoma-astrocytoma epithelial cell), whole cell lysate
Lane 5 : Hepa1-6 (mouse hepatoma epithelial cell), whole cell lysate
Lane 6 : Human testis lysate
Lane 7 : Rat testis lysate
Lane 8 : C6 (rat glial tumor glial cell), whole cell lysate
Lane 9 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate
Lane 10 : RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 36 kDa
Observed band size: 38 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure times: Lanes 1-4 & 8-9: 15 seconds; Lanes 5 & 10: 3 minutes; Lane 6: 70 seconds; Lane 7: 3 seconds.
The molecular weight observed is consistent with what has been described in the literature (PMID:23547718; PMID:25909225).
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cell line labeling PBK/SPK with ab236872 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.
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PBK/SPK was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate with ab236872 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab236872 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: HepG2 whole cell lysate 10 µg (Input).
Lane 2: ab236872 IP in HepG2 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab236872 in HepG2 whole cell lysate.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 10 seconds. -
Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling PBK/SPK with ab236872 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in germ cells of rat testis (PMID:16982762; PMID:25909225) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). -
Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling PBK/SPK with ab236872 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in germ cells of mouse testis (PMID:16982762; PMID:25909225) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). -
Immunohistochemical analysis of paraffin-embedded human gastric cancer tissue labeling PBK/SPK with ab236872 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in human gastric cancer (PMID:26894977; PMID:27898655) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). -
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cell line labeling PBK/SPK with ab236872 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.
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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SDS download
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Datasheet download
Certificate of Compliance
参考文献 (2)
ab236872 は 2 報の論文で使用されています。
- Chen SM et al. A multi-targeting natural product, aiphanol, inhibits tumor growth and metastasis. Am J Cancer Res 12:4930-4953 (2022). PubMed: 36504899
- Jiang Y et al. TOPK promotes metastasis of esophageal squamous cell carcinoma by activating the Src/GSK3ß/STAT3 signaling pathway via ?-catenin. BMC Cancer 19:1264 (2019). PubMed: 31888532