Antibodies, Proteins, Kits and Reagents for Life Science

335 040 ₸ Product size

100 µl 335 040 ₸

Order now and get it on Tuesday March 02, 2021

Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [E161] to CDK1 + Cdk2 + Cdk3 (phospho T14)
Suitable for: WB, IHC-P, IP, ICC/IF
Reacts with: Mouse, Rat, Human

Product name

Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161]
Description

Rabbit monoclonal [E161] to CDK1 + Cdk2 + Cdk3 (phospho T14)
Host species

Rabbit

Tested Applications & Species

Application	Species
ICC/IF	Human
IHC-P	Human
IP	Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Synthetic peptide within Human CDK1 aa 1-100. The exact sequence is proprietary.
Database link: P06493
Positive control
- WB: HeLa cell lysate. IHC-P: Human lymphoma.
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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

E161
Isotype

IgG
Research areas

Western blot - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)

All lanes : Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384) at 1/2000 dilution (purified)

Lane 1 : Empty vector transfected 293T (human embryonic kidney) whole cell lysates
Lane 2 : CDK1-GFP transfected 293T (human embryonic kidney) whole cell lysates
Lane 3 : CDK2-GFP transfected 293T (human embryonic kidney) whole cell lysates
Lane 4 : CDK3-GFP transfected 293T (human embryonic kidney) whole cell lysates
Lane 5 : CDK5-GFP transfected 293T (human embryonic kidney) whole cell lysates

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 34 kDa
Observed band size: 62, 34 kDa
why is the actual band size different from the predicted?

Exposure time: 15 seconds

Blocking/Diluting buffer 5% NFDM/TBST

34kDa band represents endogenous CDK, and 62kDa represents CDK-GFP.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)

Immunohistochemical staining of paraffin embedded human B cell lymphoma with purified ab32384 at a working dilution of 1/500. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Immunocytochemistry/ Immunofluorescence - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)

Immunofluorescence staining of HeLa cells with purified ab32384 at a working dilution of 1/200, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor^® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab32384 was used at a dilution of 1/500 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor^® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
Western blot - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)

All lanes : Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384) at 1/2000 dilution (purified)

Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysates
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with 3mM hydroxyurea for 20 hours whole cell lysates
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with 3mM hydroxyurea for 20 hours whole cell lysates. Then the membrane was incubated with phosphatase.

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 34 kDa
Observed band size: 34 kDa

Exposure time: 30 seconds

Blocking/Diluting buffer
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)

Immunohistochemical staining of paraffin embedded human ovarian carcinoma with purified ab32384 at a working dilution of 1/500. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Western blot - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)

Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384) at 1/5000 dilution (purified) + Raw264.7 cell lysate at 10 µg

Secondary
HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

Predicted band size: 34 kDa
Observed band size: 34 kDa

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Western blot - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)

All lanes : Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384) at 1/10000 dilution (purified)

Lane 1 : C6 cell lysate
Lane 2 : PC-12 cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

Predicted band size: 34 kDa
Observed band size: 34 kDa

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Western blot - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)

All lanes : Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384) at 1/10000 dilution (purified)

Lane 1 : A431 cell lysate
Lane 2 : HepG2 cell lysate
Lane 3 : SK-BR-3 cell lysate
Lane 4 : HeLa cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

Predicted band size: 34 kDa
Observed band size: 34 kDa

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Immunoprecipitation - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)

ab32384 (purified) at 1/50 immunoprecipitating CDK1 in 10 μg HEK293 (Lanes 1 and 2, observed at 34 kDa). Lane 3 - PBS. For western blotting, HRP Veriblot for IP Detection Reagent (ab131366) was used for detection (1/10 000). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)

Unpurified ab32384, at a 1/50 dilution, staining Cdc2 in paraffin embedded human lymphoma tissue sections by Immunohistochemistry.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Western blot - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)

All lanes : Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384) at 1/1000 dilution (unpurified)

Lane 1 : A431 cell lysate
Lane 2 : HepG2 cell lysate
Lane 3 : SKBR3 cell lysate
Lane 4 : HeLa cell lysate

Lysates/proteins at 10 µg per lane.

Predicted band size: 34 kDa
Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)

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