Recombinant Anti-Cdk8 antibody [EPR21005] (ab229192)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR21005] to Cdk8
- Suitable for: WB, IP
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Cdk8 antibody [EPR21005]
See all Cdk8 primary antibodies -
Description
Rabbit monoclonal [EPR21005] to Cdk8 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IPmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Wild-type HeLa, Daudi, HAP1, HCT 116, K562 and SW480 whole cell lysates; Human testis lysate. IP: SW480 whole cell lysate.
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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: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR21005 -
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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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab229192 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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WB |
1/1000. Detects a band of approximately 53 kDa (predicted molecular weight: 53 kDa).
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IP |
1/30.
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Notes |
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WB
1/1000. Detects a band of approximately 53 kDa (predicted molecular weight: 53 kDa). |
IP
1/30. |
Target
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Function
Component of the Mediator complex, a coactivator involved in regulated gene transcription of nearly all RNA polymerase II-dependent genes. Mediator functions as a bridge to convey information from gene-specific regulatory proteins to the basal RNA polymerase II transcription machinery. Mediator is recruited to promoters by direct interactions with regulatory proteins and serves as a scaffold for the assembly of a functional preinitiation complex with RNA polymerase II and the general transcription factors. Phosphorylates the CTD (C-terminal domain) of the large subunit of RNA polymerase II (RNAp II), which may inhibit the formation of a transcription initiation complex. Phosphorylates CCNH leading to down-regulation of the TFIIH complex and transcriptional repression. Recruited through interaction with MAML1 to hyperphosphorylate the intracellular domain of NOTCH, leading to its degradation. -
Sequence similarities
Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. CDC2/CDKX subfamily.
Contains 1 protein kinase domain. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 1024 Human
- Omim: 603184 Human
- SwissProt: P49336 Human
- Unigene: 382306 Human
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Alternative names
- cdk8 antibody
- CDK8 protein kinase antibody
- CDK8_HUMAN antibody
see all
Images
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All lanes : Anti-Cdk8 antibody [EPR21005] (ab229192) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CDK8 knockout HeLa cell lysate
Lane 3 : HAP1 cell lysate
Lane 4 : Daudi cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 53 kDaLanes 1-4: Merged signal (red and green). Green - ab229192 observed at 53 kDa. Red - loading control ab8245 observed at 37 kDa.
ab229192 Anti-Cdk8 antibody [EPR21005] was shown to specifically react with Cdk8 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265087 (knockout cell lysate ab257885) was used. Wild-type and Cdk8 knockout samples were subjected to SDS-PAGE. ab229192 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 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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Cdk8 was immunoprecipitated from 0.35 mg of SW480 (human colorectal adenocarcinoma cell line) whole cell lysate with ab229192 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab229192 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/10,000 dilution.
Lane 1: SW480 whole cell lysate 10 μg (Input).
Lane 2: ab229192 IP in SW480 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab229192 in SW480 whole cell lysate.Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes. -
All lanes : Anti-Cdk8 antibody [EPR21005] (ab229192) at 1/1000 dilution
Lane 1 : Human testis lysate
Lane 2 : K562 (human chronic myelogenous leukemia cell line from bone marrow ) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
Lane 1 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Lane 2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDa
Exposure time: 58 secondsBlocking and dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Cdk8 antibody [EPR21005] (ab229192) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate at 20 µg
Lane 2 : Cdk8 knockout HAP1 whole cell lysate at 20 µg
Lane 3 : HCT 116 (human colorectal carcinoma cell line) whole cell lysate at 10 µg
Lane 4 : SW480 (Human colorectal adenocarcinoma 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: 53 kDa
Exposure time: 48 secondsBlocking and dilution buffer: 5% NFDM/TBST.
ab229192 was shown to specifically react with Cdk8 in wild-type HAP1 cells as signal was lost in Cdk8 knockout cells.
Wild-type and Cdk8 knockout samples were subjected to SDS-PAGE. ab229192 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. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.
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
Certificate of Compliance
References (2)
ab229192 has been referenced in 2 publications.
- Dimitrova E et al. Distinct roles for CKM-Mediator in controlling Polycomb-dependent chromosomal interactions and priming genes for induction. Nat Struct Mol Biol 29:1000-1010 (2022). PubMed: 36220895
- Piao M & Zhang L Knockdown of SNHG16 suppresses the proliferation and induces the apoptosis of leukemia cells via miR-193a-5p/CDK8. Int J Mol Med 46:1175-1185 (2020). PubMed: 32705162