Antibodies, Proteins, Kits and Reagents for Life Science

338 390 ₸ Product size

100 µl 338 390 ₸

Order now and get it on Tuesday March 09, 2021

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EP569Y] to PDPK1
Suitable for: WB, IHC-P
Knockout validated
Reacts with: Mouse, Rat, Human, Pig

Product name

Anti-PDPK1 antibody [EP569Y]
See all PDPK1 primary antibodies
Description

Rabbit monoclonal [EP569Y] to PDPK1
Host species

Rabbit
Specificity

This antibody detects PDK1 both when phosphorylated and non-phosphorylated on Tyrosine 373 and 376.

The mouse, rat and pig recommendation is based on the WB results. We do not guarantee IHC-P for mouse, rat and pig.

Tested Applications & Species

Application	Species
IHC-P	Human
WB	Mouse Rat Human Pig

See all applications and species data

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding Tyr373 and Tyr376 of human PDPK1.
Positive control
- WB: PC-12, Rat testis, HEK-293, 22Rv1, PIEC, HeLa, K-562, and F9 lysates. IHC-P: human lung carcinoma and human prostate carcinoma tissue.
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. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
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

EP569Y
Isotype

IgG
Research areas

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDPK1 antibody [EP569Y] (ab52893)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue sections labeling PDPK1 with Purified ab52893 at 1:150 dilution(2.4 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Purified)ImmunoHistoProbe one step HRP Polymer (ready to use) was used for detection. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Western blot - Anti-PDPK1 antibody [EP569Y] (ab52893)

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: PDPK1 knockout HAP1 cell lysate (20 µg)
Lane 3: HEK293 cell lysate (20 µg)
Lane 4: MCF7 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab52893 observed at 70 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab52893 was shown to recognize PDPK1 when PDPK1 knockout samples were used, along with additional cross-reactive bands. Wild-type and PDPK1 knockout samples were subjected to SDS-PAGE. ab52893 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. 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 1/10000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-PDPK1 antibody [EP569Y] (ab52893)

Anti-PDPK1 antibody [EP569Y] (ab52893) at 1/1000 dilution (Purified) + F9 (Mouse embryonal carcinoma epithelial cell) whole cell lysates at 15 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/5000 dilution

Predicted band size: 63 kDa
Observed band size: 52-65 kDa
why is the actual band size different from the predicted?

The molecular weight observed is consistent with what has been described in PMID: 30171166
Western blot - Anti-PDPK1 antibody [EP569Y] (ab52893)

All lanes : Anti-PDPK1 antibody [EP569Y] (ab52893) at 1/1000 dilution (Purified)

Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/5000 dilution

Predicted band size: 63 kDa
Observed band size: 52-65 kDa why is the actual band size different from the predicted?

The molecular weight observed is consistent with what has been described in PMID: 30171166
Western blot - Anti-PDPK1 antibody [EP569Y] (ab52893)

All lanes : Anti-PDPK1 antibody [EP569Y] (ab52893) at 1/1000 dilution (Purified)

Lane 1 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates
Lane 2 : Rat testis lysates
Lane 3 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates
Lane 4 : 22Rv1 (Human prostate carcinoma epithelial cell) whole cell lysates
Lane 5 : PIEC (PIG iliac artery endothelial cell) whole cell lysates

Lysates/proteins at 20 µg per lane.

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

Predicted band size: 63 kDa
Observed band size: 52-65 kDa why is the actual band size different from the predicted?

The molecular weight observed is consistent with what has been described in PMID: 30171166
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDPK1 antibody [EP569Y] (ab52893)

Ab52893 (1:100) staining human PDPK1 in human prostate carcinoma tissue by immunohistochemistry using paraffin embedded tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Western blot - Anti-PDPK1 antibody [EP569Y] (ab52893)

Anti-PDPK1 antibody [EP569Y] (ab52893) at 1/1000 dilution + 293 cell lysate

Predicted band size: 63 kDa
Observed band size: 63 kDa
Anti-PDPK1 antibody [EP569Y] (ab52893)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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