Recombinant Anti-PBK/SPK antibody [EPR21983] KO Tested (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

Product name

Anti-PBK/SPK antibody [EPR21983]
See all PBK/SPK primary antibodies
Description

Rabbit monoclonal [EPR21983] to PBK/SPK
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), WB, IHC-P, IPmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- 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.
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
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.

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

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR21983
Isotype

IgG
Research areas

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

Our Abpromise guarantee covers the use of ab236872 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
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.

Notes
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.

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
modifications

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
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
Alternative names
- 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

SDS download

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Datasheet download

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Publishing research using ab236872? Please let us know so that we can cite the reference in this datasheet.

ab236872 has been referenced in 2 publications.

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

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

ChIP Related Products

Compatible Secondaries

Immunohistochemistry reagents

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Post-translationalmodifications

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (2)

Customer reviews and Q&As

Post-translational
modifications