Anti-Cdk8 antibody [EPR26566-270]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk8 antibody [EPR26566-270] (AB325133)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Cdk8 with ab325133 at 1/500 (1.008 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human colon.
The primary antibody was incubated 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 a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk8 antibody [EPR26566-270] (AB325133)

Immunohistochemical analysis of paraffin-embedded (A) Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) cell pellet and (B) CDK8 knockout HAP1 cell pellet labeling Cdk8 with ab325133 at 1/1000 (0.504 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in (A) Wild-type HAP1 cell pellet, and no signal in (B) CDK8 knockout HAP1 cell pellet.
The primary antibody was incubated 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 a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk8 antibody [EPR26566-270] (AB325133)

Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labeling Cdk8 with ab325133 at 1/500 (1.008 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human colon cancer.
The primary antibody was incubated 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 a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• WB

Supplier Data

Western blot - Anti-Cdk8 antibody [EPR26566-270] (AB325133)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, ab325133 was shown to bind specifically to Cdk8. Target of interest was observed at 53 kDa in wild-type HAP1 cell lysates (lane

1. with no signal observed at this size in Cdk8 knockout cell line (lane 2).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 32596239; PMID : 23746844).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Cdk8 antibody [EPR26566-270] (ab325133) at 1/1000 dilution

Lane 1:

Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell ) whole cell lysate at 20 µg

Lane 2:

CDK8 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

SW480 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

Daudi (human Burkitt's lymphoma lymphoblast) whole cell lysate at 20 µg

Lane 6:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 7:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 53 kDa,36 kDa

false

Exposure time: 81s

• WB

Supplier Data

Western blot - Anti-Cdk8 antibody [EPR26566-270] (AB325133)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 32596239; PMID : 23746844).

The identity of the lower MW band at approximately 30 kDa is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Cdk8 antibody [EPR26566-270] (ab325133) at 1/1000 dilution

Lane 1:

Human colon tissue lysate at 48 µg

Lane 2:

Human colon cancer tissue lysate at 48 µg

Lane 3:

Human pancreas tissue lysate at 48 µg

Lane 4:

Human pancreas cancer tissue lysate at 48 µg

Lane 5:

Human testis tissue lysate at 48 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 53 kDa,36 kDa

false

Exposure time: 103s

• I-ELISA

Supplier Data

Indirect ELISA - Anti-Cdk8 antibody [EPR26566-270] (AB325133)

Indirect ELISA analysis of ab325133 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution dilution.

Antigen : Human CDK8, Human CDK19.

Antigen concentration : 1000 ng/ml

This antibody does not cross-react with human CDK19