Antibodies, Proteins, Kits and Reagents for Life Science

304 886 ₸ Product size

100 µl 304 886 ₸

Order now and get it on Tuesday March 02, 2021

Anti-PKA R2/PKR2 antibody [Y116] (ab32514)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [Y116] to PKA R2/PKR2
Suitable for: ICC, IP, WB, IHC-P, Flow Cyt
Knockout validated
Reacts with: Human

Product name

Anti-PKA R2/PKR2 antibody [Y116]
See all PKA R2/PKR2 primary antibodies
Description

Rabbit monoclonal [Y116] to PKA R2/PKR2
Host species

Rabbit
Specificity

The antibody recognises PKA RII. It does not cross-react with other cAMP-dependent kinase.

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC	Human
IHC-P	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Synthetic peptide within Human PKA R2/PKR2 aa 350-450 (C terminal). The exact sequence is proprietary.
Positive control
- WB: K562, HeLa and Jurkat cell lysates. IHC-P: Human colon cancer tissue. IP: K562 whole cell lysate. Flow Cyt: MCF7 cells. ICC: HeLa cells.
General notes
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

This product was previously labelled as PKA R2.
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 long term. 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

Y116
Isotype

IgG
Research areas

Western blot - Anti-PKA R2/PKR2 antibody [Y116] (ab32514)

All lanes : Anti-PKA R2/PKR2 antibody [Y116] (ab32514) at 1/10000 dilution (Purified)

Lane 1 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate

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: 45 kDa
Observed band size: 50 kDa
why is the actual band size different from the predicted?
Immunoprecipitation - Anti-PKA R2/PKR2 antibody [Y116] (ab32514)

Purified ab32514 at 1/20 dilution (0.5µg) immunoprecipitating PKA R2/PKR2 in K-562 whole cell lysate.
Lane 1 (input): K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate 10µg
Lane 2 (+): ab32514 + K-562 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32514 in K-562 whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 50 kDa
Flow Cytometry - Anti-PKA R2/PKR2 antibody [Y116] (ab32514)

Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling PKA R2/PKR2 with purified ab32514 at 1/20 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
Immunocytochemistry - Anti-PKA R2/PKR2 antibody [Y116] (ab32514)

Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling PKA R2/PKR2 with purified ab32514 at 1/500 dilution (1.9 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKA R2/PKR2 antibody [Y116] (ab32514)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon cancer tissue sections labeling PKA R2/PKR2 with purified ab32514 at 1/100 dilution (0.84 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Western blot - Anti-PKA R2/PKR2 antibody [Y116] (ab32514)

All lanes : Anti-PKA R2/PKR2 antibody [Y116] (ab32514) at 1/1000 dilution (Unpurified)

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : PRKAR2A knockout HeLa cell lysate
Lane 3 : K-562 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 45 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?

Lanes 1-3: Merged signal (red and green). Green - ab32514 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa.

Unpurified ab32514 Anti-PKA R2/PKR2 antibody [Y116] was shown to specifically react with PKA R2/PKR2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265748 (knockout cell lysate ab257607) was used. Wild-type and PKA R2/PKR2 knockout samples were subjected to SDS-PAGE. ab32514 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours 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.
Anti-PKA R2/PKR2 antibody [Y116] (ab32514)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com