Recombinant Anti-PBK/SPK antibody [EP2520Y] (ab75987)
![Western blot - Anti-PBK/SPK antibody [EP2520Y] (ab75987)](https://www.abcam.com/ps/products/75/ab75987/Images/ab75987-259084-anti-pbk-antibody-ep2520y-western-blot.jpg "Western blot - Anti-PBK/SPK antibody [EP2520Y] (ab75987)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP2520Y] to PBK/SPK
- Suitable for: WB, IP, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-PBK/SPK antibody [EP2520Y]
See all PBK/SPK primary antibodies -
Description
Rabbit monoclonal [EP2520Y] to PBK/SPK -
Host species
Rabbit -
Tested applications
Suitable for: WB, IP, IHC-Pmore details
Unsuitable for: Flow Cyt or ICC -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human PBK/SPK (C terminal). The exact sequence is proprietary.
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Positive control
- WB: HEK-293T, HeLa and HAP1 cell lysates. IHC-P: Human tonsil tissue
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General notes
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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP2520Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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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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KO cell pellets
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab75987 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB |
1/100000 - 1/500000. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).
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| IP |
1/30.
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| IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Antigen retrieval is recommended. |
| Notes |
|---|
|
WB
1/100000 - 1/500000. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa). |
|
IP
1/30. |
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IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Antigen retrieval is recommended. |
Target
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Function
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. -
Tissue specificity
Expressed in the testis and placenta. In the testis, restrictedly expressed in outer cell layer of seminiferous tubules. -
Sequence similarities
Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. MAP kinase kinase subfamily.
Contains 1 protein kinase domain. -
Post-translational
modificationsPhosphorylated; in a cell-cycle dependent manner at mitosis. - Information by UniProt
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Database links
- Entrez Gene: 55872 Human
- Omim: 611210 Human
- SwissProt: Q96KB5 Human
- Unigene: 104741 Human
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Alternative names
- Cancer/testis antigen 84 antibody
- CT84 antibody
- Epididymis luminal protein 164 antibody
see all
Images
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Western blot - Anti-PBK/SPK antibody [EP2520Y] (ab75987)
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: PBK knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: Human tonsil tissue lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab75987 observed at 40 kDa. Red - loading control, ab7291, observed at 52 kDa.
ab75987 was shown to recognize PBK when PBK knockout samples were used, along with additional cross-reactive bands. Wild-type and PBK knockout samples were subjected to SDS-PAGE. ab75987 diluted to 1/100,000 and ab7291 (loading control to alpha Tubulin) diluted to 1/10,000 and incubated overnight at 4°C. 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 1/10,000 dilution for 1 hour at room temperature before imaging. -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBK/SPK antibody [EP2520Y] (ab75987)](https://www.abcam.com/ps/products/75/ab75987/Images/ab75987-91235-anti-pbk-antibody-ep2520y-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBK/SPK antibody [EP2520Y] (ab75987)Immunohistochemical analysis of paraffin-embedded human tonsil using ab75987 at 1/100 dilution. Antigen retrieval was heat mediated with citrate buffer pH 6 before commencing with IHC staining protocol.
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![Western blot - Anti-PBK/SPK antibody [EP2520Y] (ab75987)](https://www.abcam.com/ps/products/75/ab75987/Images/ab75987-409561-anti-pbkspk-antibody-ep2520y-western-blot-hek-293t-lysates.jpg)
Western blot - Anti-PBK/SPK antibody [EP2520Y] (ab75987)All lanes : Anti-PBK/SPK antibody [EP2520Y] (ab75987) at 1/100000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : PBK knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 36 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab75987 observed at 40 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab75987 was shown to react with PBK/SPK in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266827 (knockout cell lysate ab257575) was used. Wild-type HEK-293T and PBK knockout HEK-293T cell lysates were subjected to SDS-PAGE. ab75987 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 100000 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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![Anti-PBK/SPK antibody [EP2520Y] (ab75987)](https://www.abcam.com/ps/products/75/ab75987/Images/ab75987-7-benefits-of-recombinant-antibodies.png)
Anti-PBK/SPK antibody [EP2520Y] (ab75987)
Protocols
Datasheets and documents
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Datasheet download
References (7)
ab75987 has been referenced in 7 publications.
- Mao P et al. PDZ-Binding Kinase-Dependent Transcriptional Regulation of CCNB2 Promotes Tumorigenesis and Radio-Resistance in Glioblastoma. Transl Oncol 13:287-294 (2020). PubMed: 31874375
- Park JH et al. TOPK inhibition accelerates oxidative stress-induced granulosa cell apoptosis via the p53/SIRT1 axis. Int J Mol Med 46:1923-1937 (2020). PubMed: 32901866
- Ma H et al. PBK, targeted by EVI1, promotes metastasis and confers cisplatin resistance through inducing autophagy in high-grade serous ovarian carcinoma. Cell Death Dis 10:166 (2019). PubMed: 30778048
- Huo X et al. Clinical and Expression Significance of AKT1 by Co-expression Network Analysis in Endometrial Cancer. Front Oncol 9:1147 (2019). PubMed: 31781484
- Dou X et al. PBK/TOPK mediates geranylgeranylation signaling for breast cancer cell proliferation. Cancer Cell Int 15:27 (2015). Human . PubMed: 25745361
- Lei B et al. PBK/TOPK expression correlates with mutant p53 and affects patients' prognosis and cell proliferation and viability in lung adenocarcinoma. Hum Pathol 46:217-24 (2015). PubMed: 25466965
- Guo Z et al. Proteomics strategy to identify substrates of LNX, a PDZ domain-containing E3 ubiquitin ligase. J Proteome Res 11:4847-62 (2012). PubMed: 22889411
