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RecombinantRabMAb

Recombinant Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] - BSA and Azide free (ab227848)

Submit a review Submit a question References (7)
Western blot - Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] - BSA and Azide free (ab227848)
  • Western blot - Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] - BSA and Azide free (ab227848)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] - BSA and Azide free (ab227848)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] - BSA and Azide free (ab227848)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] - BSA and Azide free (ab227848)
  • Dot Blot - Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] - BSA and Azide free (ab227848)
  • Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] - BSA and Azide free (ab227848)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP2606Y] to PKA alpha/beta/gamma (catalytic subunit) (phospho T197) - BSA and Azide free
  • Suitable for: WB, IHC-P, Dot blot
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Unconjugated

Overview

  • Product name

    Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] - BSA and Azide free
    See all PKA alpha/beta/gamma (catalytic subunit) primary antibodies

  • Description

    Rabbit monoclonal [EP2606Y] to PKA alpha/beta/gamma (catalytic subunit) (phospho T197) - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, Dot blotmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: NIH/3T3, C6, HeLa cell lysate treated with calyculin A IHC: human liver carcinoma, mouse kidney, rat kidney

  • General notes

    ab227848 is the carrier-free version of ab75991.

    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.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab227848 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB
Use at an assay dependent concentration. Predicted molecular weight: 40 kDa.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Use of HRP-conjugated or polymerized HRP secondary antibodies recommended, stronger signals have been found using the polymerized HRP secondary.
Dot blot
Use at an assay dependent concentration.
Notes
WB
Use at an assay dependent concentration. Predicted molecular weight: 40 kDa.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Use of HRP-conjugated or polymerized HRP secondary antibodies recommended, stronger signals have been found using the polymerized HRP secondary.
Dot blot
Use at an assay dependent concentration.

Target

Images

  • Western blot - Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] - BSA and Azide free (ab227848)
    Western blot - Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] - BSA and Azide free (ab227848)
    All lanes : Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] (ab75991) at 1/1000 dilution (Purified)

    Lane 1 : Untreated HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) starved overnight, then treated with 100nM Calyculin A for 30 minutes, whole cell lysate
    Lane 3 : HeLa (Human cervix adenocarcinoma epithelial cell) starved overnight, then treated with 100nM Calyculin A for 30 minutes whole cell lysate, then the membrane treated with Lambda Phosphatase for 1 hour

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

    Predicted band size: 40 kDa
    Observed band size: 40 kDa



    This data was developed using ab75991, the same antibody clone in a different buffer formulation.

  • Western blot - Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] - BSA and Azide free (ab227848)
    Western blot - Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] - BSA and Azide free (ab227848)
    All lanes : Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] (ab75991) at 1/1000 dilution (Purified)

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate, then the membrane treated with Lambda Phosphatase for 1 hour
    Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
    Lane 4 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate, then the membrane treated with Lambda Phosphatase for 1 hour
    Lane 5 : C6 (Rat glial tumor glial cell) whole cell lysate
    Lane 6 : C6 (Rat glial tumor glial cell) whole cell lysate, then the membrane treated with Lambda Phosphatase for 1 hour

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

    Predicted band size: 40 kDa
    Observed band size: 40 kDa



    This data was developed using ab75991, the same antibody clone in a different buffer formulation.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] - BSA and Azide free (ab227848)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] - BSA and Azide free (ab227848)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue sections labeling PKA alpha/beta/gamma (catalytic subunit) with Purified ab75991 at 1/5000 dilution (0.05 µg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control. M3

    This data was developed using ab75991, the same antibody clone in a different buffer formulation.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] - BSA and Azide free (ab227848)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] - BSA and Azide free (ab227848)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling PKA alpha/beta/gamma (catalytic subunit) with Purified ab75991 at 1/5000 dilution (0.05 µg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control. M3

    This data was developed using ab75991, the same antibody clone in a different buffer formulation.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] - BSA and Azide free (ab227848)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] - BSA and Azide free (ab227848)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver carcinoma tissue sections labeling PKA alpha/beta/gamma (catalytic subunit) with Purified ab75991 at 1/5000 dilution (0.05 µg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control. M3

    This data was developed using ab75991, the same antibody clone in a different buffer formulation.

  • Dot Blot - Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] - BSA and Azide free (ab227848)
    Dot Blot - Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] - BSA and Azide free (ab227848)

    Dot blot analysis of various peptides labelling PKA gamma (catalytic subunit) (phospho T197) with ab75991 at a dilution of 1/1000. ab97051, a peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/100000).

    Lane 1 - PKA alpha/beta/gamma (catalytic subunit) (pT197) phospho peptide.

    Lane 2 - PKA alpha/beta/gamma (catalytic subunit) non-phospho peptide.

    Lane 3 - PRKX/Y (pT203) phospho peptide.

    Lane 4 - PRKX/Y non-phospho peptide.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75991).

  • Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] - BSA and Azide free (ab227848)
    Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] - BSA and Azide free (ab227848)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (7)

Publishing research using ab227848? Please let us know so that we can cite the reference in this datasheet.

ab227848 has been referenced in 7 publications.

  • Ni S  et al. Sensory innervation in porous endplates by Netrin-1 from osteoclasts mediates PGE2-induced spinal hypersensitivity in mice. Nat Commun 10:5643 (2019). PubMed: 31822662
  • Li J  et al. Linarin promotes osteogenic differentiation by activating the BMP-2/RUNX2 pathway via protein kinase A signaling. Int J Mol Med 37:901-10 (2016). PubMed: 26935542
  • Ferdaus MZ  et al. SPAK and OSR1 play essential roles in potassium homeostasis through actions on the distal convoluted tubule. J Physiol 594:4945-66 (2016). PubMed: 27068441
  • Han YS  et al. Human heart failure is accompanied by altered protein kinase A subunit expression and post-translational state. Arch Biochem Biophys 538:25-33 (2013). PubMed: 23942052
  • Fonseca SG  et al. Wolfram syndrome 1 and adenylyl cyclase 8 interact at the plasma membrane to regulate insulin production and secretion. Nat Cell Biol 14:1105-12 (2012). PubMed: 22983116
  • Li Y  et al. STAT1, STAT6 and adenosine 3',5'-cyclic monophosphate (cAMP) signaling drive SOCS3 expression in inactive ulcerative colitis. Mol Med 18:1412-9 (2012). PubMed: 23154639
  • Niculescu LS  et al. Apolipoprotein A-I stimulates cholesteryl ester transfer protein and apolipoprotein E secretion from lipid-loaded macrophages; the role of NF-?B and PKA signaling pathways. Biochem Biophys Res Commun 415:497-502 (2011). WB ; Human . PubMed: 22056562

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