Recombinant Anti-ZIP Kinase antibody [EPR18809-86] (ab210528)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18809-86] to ZIP Kinase
- Suitable for: Flow Cyt (Intra), WB, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human, Recombinant fragment
Related conjugates and formulations
Overview
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Product name
Anti-ZIP Kinase antibody [EPR18809-86]
See all ZIP Kinase primary antibodies -
Description
Rabbit monoclonal [EPR18809-86] to ZIP Kinase -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human, Recombinant fragment -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human ZIP Kinase fragment recombinant protein; Human fetal liver, fetal heart, fetal kidney and bladder lysates; Jurkat, A431, HeLa, C2C12, L6, A549, C6, RAW 264.7, PC-12 and NIH/3T3 cell lysates; Mouse and rat bladder lysates; Mouse brain, kidney and spleen lysates; Rat brain, heart, kidney and spleen lysates. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt (intra): C2C12 and NIH/3T3 cells.
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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.
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
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18809-86 -
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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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab210528 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) |
1/120.
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WB |
1/1000. Detects a band of approximately 50 kDa (predicted molecular weight: 53 kDa).
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ICC/IF |
1/100.
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Notes |
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Flow Cyt (Intra)
1/120. |
WB
1/1000. Detects a band of approximately 50 kDa (predicted molecular weight: 53 kDa). |
ICC/IF
1/100. |
Target
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Function
Serine/threonine kinase which acts as a positive regulator of apoptosis. Phosphorylates histone H3 on 'Thr-11' at centromeres during mitosis. Regulates myosin light chain phosphatase through phosphorylation of MYPT1 thereby regulating the assembly of the actin cytoskeleton, cell migration, invasiveness of tumor cells, smooth muscle contraction and neurite outgrowth. Involved in the formation of promyelocytic leukemia protein nuclear body (PML-NB), one of many subnuclear domains in the eukaryotic cell nucleus, and which is involved in oncogenesis and viral infection. -
Sequence similarities
Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. DAP kinase subfamily.
Contains 1 protein kinase domain. -
Post-translational
modificationsUbiquitinated. Ubiquitination mediated by the UBE2D3 E3 ligase does not lead to proteasomal degradation, but influences promyelocytic leukemia protein nuclear bodies (PML-NBs) formation in the nucleus.
Autophosphorylated. Phosphorylated by ROCK1. -
Cellular localization
Nucleus. Cytoplasm. Nucleus > PML body. Relocates to the cytoplasm on binding PAWR where the complex appears to interact with actin filaments (By similarity). Localizes to promyelocytic leukemia protein nuclear bodies (PML-NBs). Associates to centromeres from prophase to anaphase. - Information by UniProt
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Database links
- Entrez Gene: 1613 Human
- Entrez Gene: 13144 Mouse
- Entrez Gene: 64391 Rat
- Omim: 603289 Human
- SwissProt: O43293 Human
- SwissProt: O54784 Mouse
- SwissProt: O88764 Rat
- Unigene: 631844 Human
see all -
Alternative names
- DAP kinase 3 antibody
- DAP like kinase antibody
- DAP-like kinase antibody
see all
Images
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Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: ZIP Kinase knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: A431 whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab210528 observed at 53 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab210528 was shown to specifically react with ZIP Kinase in wild type cells as signal was lost in ZIP Kinase knockout cells. Wild-type and ZIP Kinase knockout samples were subjected to SDS-PAGE. Ab210528 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at a 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-ZIP Kinase antibody [EPR18809-86] (ab210528) at 1/1000 dilution
Lane 1 : Human ZIP Kinase fragment recombinant protein
Lane 2 : Human DAP Kinase 2 fragment recombinant protein
Lane 3 : Human DAP Kinase 1 fragment recombinant protein
Lysates/proteins at 0.01 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 53 kDa
Observed band size: 29 kDa why is the actual band size different from the predicted?
Exposure time: 1 secondBlocking/Dilution buffer: 5% NFDM/TBST.
Human ZIP Kinase fragment recombinant protein contain aa13-275 with a His-Tag®. Human DAP Kinase 2 fragment recombinant protein contain aa23-285 with a His-Tag®. Human DAP Kinase 1 fragment recombinant protein contain aa13-275 with a His-Tag®. All three recombinant human fragment proteins were made in-house.
The antibody reacts weakly with DAP kinase 1 and DAP kinase 2; however bands of their appropriate MW (159 kD, 42 kD) are not detected in the tissue lysates.
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All lanes : Anti-ZIP Kinase antibody [EPR18809-86] (ab210528) at 1/1000 dilution
Lane 1 : Human fetal liver lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate
Lane 4 : Human bladder lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 53 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID: 20124481).
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All lanes : Anti-ZIP Kinase antibody [EPR18809-86] (ab210528) at 1/1000 dilution
Lane 1 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 2 : A431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 4 : C2C12 (Mouse myoblast cell line) whole cell lysate
Lane 5 : L6 (Rat skeletal muscle cell line) whole cell lysate
Lane 6 : Mouse bladder lysate
Lane 7 : Rat bladder lysate
Lane 8 : A549 (Human lung carcinoma cell line) 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: 53 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1, 2, 3, 4, 5, 6 and 7: 30 seconds; Lane 8: 5 seconds.
The molecular weight observed is consistent with what has been described in the literature (PMID: 20124481).
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All lanes : Anti-ZIP Kinase antibody [EPR18809-86] (ab210528) at 1/1000 dilution
Lane 1 : Mouse brain tissue lysate at 10 µg
Lane 2 : Mouse kidney tissue lysate at 10 µg
Lane 3 : Mouse spleen tissue lysate at 10 µg
Lane 4 : Rat brain tissue lysate at 10 µg
Lane 5 : Rat heart tissue lysate at 10 µg
Lane 6 : Rat kidney tissue lysate at 10 µg
Lane 7 : Rat spleen tissue lysate at 10 µg
Lane 8 : C6 (Rat glial tumor cell line) whole cell lysate at 20 µg
Lane 9 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg
Lane 10 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg
Lane 11 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 53 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1, 2 and 3: 30 seconds; Lane 4, 5, 6 and 7: 3 minutes; Lane 8, 9, 10 and 11: 30 seconds.
The molecular weight observed is consistent with what has been described in the literature (PMID: 20124481)
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling ZIP Kinase with ab210528 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasm and nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab210528 at 1/100 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution. -
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling ZIP Kinase with ab210528 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasm and nuclear staining on NIH/3T3 cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab210528 at 1/100 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution. -
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed C2C12 (Mouse myoblast cell line) cells labeling ZIP Kinase with ab210528 at 1/120 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor® 488) at 1/500 dilution was used as the secondary antibody.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling ZIP Kinase with ab210528 at 1/600 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor® 488) at 1/500 dilution was used as the secondary antibody.
Note:Cells were permeabilised with 90% methanol -1XPBS (-20?, 30min).
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (1)
ab210528 has been referenced in 1 publication.
- Kennedy MA et al. TRANSPIRE: A Computational Pipeline to Elucidate Intracellular Protein Movements from Spatial Proteomics Data Sets. J Am Soc Mass Spectrom 31:1422-1439 (2020). PubMed: 32401031