Recombinant Anti-PBK/SPK antibody [31/PBK] (ab280209)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Mouse monoclonal [31/PBK] to PBK/SPK
- Suitable for: Flow Cyt (Intra), WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-PBK/SPK antibody [31/PBK]
See all PBK/SPK primary antibodies -
Description
Mouse monoclonal [31/PBK] to PBK/SPK -
Host species
Mouse -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-Pmore details
Unsuitable for: ICC/IF -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Wild-type HEK-293T whole cell lysate. HeLa, HepG2, RAW264.7, C6, PC-12 and U-87 MG whole cell lysate. Human, rat and mouse testis tissue lysate. IHC-P: Human tonsil tissue. Mouse and rat testis tissue. Flow Cyt (intra) (intra): HEK-239T cells.
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General notes
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
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
Properties
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Form
Liquid -
Storage instructions
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. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
31/PBK -
Isotype
IgG2b -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab280209 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 |
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Flow Cyt (Intra) |
1/1000.
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WB |
1/1000. Predicted molecular weight: 36 kDa.
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IHC-P |
1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Notes |
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Flow Cyt (Intra)
1/1000. |
WB
1/1000. Predicted molecular weight: 36 kDa. |
IHC-P
1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
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
- 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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Alternative names
- Cancer/testis antigen 84 antibody
- CT84 antibody
- Epididymis luminal protein 164 antibody
see all
Images
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All lanes : Anti-PBK/SPK antibody [31/PBK] (ab280209) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T (human embryonic kidney epithelial cell), whole cell lysate
Lane 2 : PBK knockout HEK-293T (human embryonic kidney epithelial cell), whole cell lysate
Lane 3 : HeLa (human epithelial cell line from cervix adenocarcinoma), whole cell lysate
Lane 4 : U-87 MG (human glioblastoma-astrocytoma epithelial cell), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Mouse IgG H&L (IRDye® 800CW) (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) (ab216777) at 1/10000 dilution
Predicted band size: 36 kDaLanes 1 - 4: Merged signal (red and green). Green - ab280209 observed at 36 kDa. Red - loading control ab52901(Rabbit monoclonal [EP1331Y] to beta Tubulin) observed at 52 kDa.
Lanes 1-2: ab280209 Anti-PBK antibody was shown to react with in HEK-293T cells in Western blot. Loss of signal was observed when PBK knockout sample was used. Wild-type and PBK knockout samples were subjected to SDS-PAGE.
ab280209 and Anti-beta Tubulin antibody [EP1331Y] (ab52901) were incubated at 4° overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680CW) (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800RD) (ab216772) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.Blocking/Dilution buffer: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.
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Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling PBK/PSK with ab280209 at 1/500 (1.946 µg/ml) dilution followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection).
Positive staining in germinal center of human tonsil.
The section was incubated with ab280209 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
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Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling PBK/SPK with ab280209 at 1/1000 dilution (0.1 µg) (red) compared with a mouse monoclonal IgG isotype control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
A Goat anti mouse IgG (Alexa Fluor® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.
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All lanes : Anti-PBK/SPK antibody [31/PBK] (ab280209) at 1/1000 dilution
Lane 1 : Human testis tissue lysate
Lane 2 : Human heart tissue lysate
Lane 3 : Mouse testis tissue lysate
Lane 4 : Mouse brain tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution
Predicted band size: 36 kDaNegative control: Human heart, mouse brain (PMID: 27898655).
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:23547718).
Blocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-PBK/SPK antibody [31/PBK] (ab280209) at 1/1000 dilution
Lane 1 : HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate
Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma), whole cell lysate
Lane 3 : U-87 MG (human glioblastoma-astrocytoma epithelial cell), whole cell lysate
Lane 4 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate
Lane 5 : C6 (rat glial tumor glial cell), whole cell lysate
Lane 6 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate
Lane 7 : Rat testis tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution
Predicted band size: 36 kDaThe expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:23547718).
Exposure time: lane 1: 37 secs; lane 2-7: 15 secs.
Blocking/Dilution buffer: 5% NFDM/TBST.
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Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling PBK/PSK with ab280209 at 1/500 (1.946 µg/ml) dilution followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection).
Positive staining in mouse testis.
The section was incubated with ab280209 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
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Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling PBK/PSK with ab280209 at 1/500 (1.946 µg/ml) dilution followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection).
Positive staining in rat testis.
The section was incubated with ab280209 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
Protocols
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.
Datasheets and documents
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SDS download
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Datasheet download
References (1)
ab280209 has been referenced in 1 publication.
- Li J et al. TOPK Affects Autophagy of Skin Squamous Cell Carcinoma by Regulating NF-KB Pathway through HDAC1. Dis Markers 2022:3771711 (2022). PubMed: 35756488