Antibodies, Proteins, Kits and Reagents for Life Science

321 638 ₸ Product size

100 µl 321 638 ₸

Order now and get it on Tuesday March 09, 2021

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

Product name

Anti-Cdk8 antibody [EPR21005]
See all Cdk8 primary antibodies
Description

Rabbit monoclonal [EPR21005] to Cdk8
Host species

Rabbit
Tested Applications & Species

Application Species
IP
Human

WB
Human

See all applications and species data
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Wild-type HeLa, Daudi, HAP1, HCT 116, K562 and SW480 whole cell lysates; Human testis lysate. IP: SW480 whole cell lysate.
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.

Application	Species
IP	Human
WB	Human

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

EPR21005
Isotype

IgG
Research areas

Western blot - Anti-Cdk8 antibody [EPR21005] (ab229192)

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 kDa

Lanes 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.
Immunoprecipitation - Anti-Cdk8 antibody [EPR21005] (ab229192)

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.
Western blot - Anti-Cdk8 antibody [EPR21005] (ab229192)

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 seconds

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-Cdk8 antibody [EPR21005] (ab229192)

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 seconds

Blocking 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.
Anti-Cdk8 antibody [EPR21005] (ab229192)

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