Anti-PBK/SPK 抗体 [EPR21983] (ab236872)

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Western blot - Anti-PBK/SPK antibody [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

Carrier Free

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.
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.
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
For more information see here.
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...
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

Alternative Versions
- Anti-PBK/SPK antibody [EPR21983] - BSA and Azide free (ab239760)
ChIP Related Products
- Anti-PBK/SPK antibody [EPR21983] - BSA and Azide free (ab239760)
Compatible Secondaries
Immunohistochemistry reagents
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant human PBK/SPK protein (ab85275)
Related Products
- Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

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
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.

特記事項
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.

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.
Target information above from: UniProt accession Q96KB5 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
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
別名
- Cancer/testis antigen 84 antibody
- CT84 antibody
- Epididymis luminal protein 164 antibody
- FLJ14385 antibody
- HEL164 antibody
- Lymphokine activated killer T cell originated protein kinase antibody
- Lymphokine-activated killer T-cell-originated protein kinase antibody
- MAPKK like protein kinase antibody
- MAPKK-like protein kinase antibody
- Nori 3 antibody
- Nori-3 antibody
- Nori3 antibody
- PBK antibody
- PDZ binding kinase antibody
- PDZ-binding kinase antibody
- Serine/threonine protein kinase antibody
- Spermatogenesis related protein kinase antibody
- Spermatogenesis-related protein kinase antibody
- SPK antibody
- T LAK cell originated protein kinase antibody
- T-LAK cell-originated protein kinase antibody
- TOPK antibody
- TOPK_HUMAN antibody
see all

Western blot - Anti-PBK/SPK antibody [EPR21983] (ab236872)

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 seconds

Blocking/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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBK/SPK antibody [EPR21983] (ab236872)

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).
Western blot - Anti-PBK/SPK antibody [EPR21983] (ab236872)

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).
Flow Cytometry (Intracellular) - Anti-PBK/SPK antibody [EPR21983] (ab236872)

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.
Immunoprecipitation - Anti-PBK/SPK antibody [EPR21983] (ab236872)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBK/SPK antibody [EPR21983] (ab236872)

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).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBK/SPK antibody [EPR21983] (ab236872)

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).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBK/SPK antibody [EPR21983] (ab236872)

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).
Flow Cytometry (Intracellular) - Anti-PBK/SPK antibody [EPR21983] (ab236872)

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.
Anti-PBK/SPK antibody [EPR21983] (ab236872)

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.

Click here to view the general protocols

Datasheet download

Download

ab236872 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab236872 は 1 報の論文で使用されています。

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

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Key features and details

Related conjugates and formulations

製品の概要

製品名

製品の詳細

由来種

アプリケーション

種交差性

免疫原

ポジティブ･コントロール

特記事項

製品の特性

製品の状態

保存方法

バッファー

精製度

ポリ/モノ

クローン名

アイソタイプ

研究分野

関連製品

Alternative Versions

ChIP Related Products

Compatible Secondaries

Immunohistochemistry reagents

Isotype control

Recombinant Protein

Related Products

アプリケーション

The Abpromise guarantee

ターゲット情報

機能

組織特異性

配列類似性

翻訳後修飾

参照データベース

別名

画像

プロトコール

データシートおよび資料

Datasheet download

Certificate of Compliance

参考文献 (1)

レビューと Q&A