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    products/primary-antibodies/pp-t-antibody-12f7-ab223367.pdf

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Validated using a knockout cell line

Anti-PP-T antibody [12F7] (ab223367)

  • Datasheet
  • SDS
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Western blot - Anti-PP-T antibody [12F7] (ab223367)
  • Immunocytochemistry/ Immunofluorescence - Anti-PP-T antibody [12F7] (ab223367)
  • Western blot - Anti-PP-T antibody [12F7] (ab223367)
  • Immunocytochemistry/ Immunofluorescence - Anti-PP-T antibody [12F7] (ab223367)

Key features and details

  • Mouse monoclonal [12F7] to PP-T
  • Suitable for: ICC/IF, WB
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG1

You may also be interested in

Primary
Product image
Anti-NXF1 antibody [EPR8009] (ab129160)
Secondary
Product image
Goat Anti-Mouse IgG H&L (HRP) (ab205719)
Knockout
Product image
Human PPP5C (PP-T) knockout HEK-293T cell line (ab266507)

View more associated products

Overview

  • Product name

    Anti-PP-T antibody [12F7]
    See all PP-T primary antibodies
  • Description

    Mouse monoclonal [12F7] to PP-T
  • Host species

    Mouse
  • Tested applications

    Suitable for: ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Recombinant full length protein corresponding to Human PP-T aa 1 to the C-terminus.
    Database link: P53041

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control

    • WB: A431, HEK-293 and Jurkat whole cell lysate (ab7899). ICC/IF: HeLa and HEK-293 cells.
  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • 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

    Preservative: 0.09% Sodium azide
    Constituents: 50% Glycerol (glycerin, glycerine), PBS
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Monoclonal
  • Clone number

    12F7
  • Isotype

    IgG1
  • Research areas

    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • RNA Processing
    • Other
    • Epigenetics and Nuclear Signaling
    • Cell cycle
    • Kinases/Phosphatases
    • Phospatases

Associated products

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
  • KO cell lines

    • Human PPP5C (PP-T) knockout HEK-293T cell line (ab266507)
  • KO cell lysates

    • Human PPP5C (PP-T) knockout HEK-293T cell lysate (ab258136)
  • Positive Controls

    • Jurkat whole cell lysate (ab30128)
    • A-431 whole cell lysate (ab30132)
    • Jurkat whole cell lysate (ab7899)
    • HEK-293 whole cell lysate (ab7902)
    • A-431 whole cell lysate (ab7909)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab223367 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
ICC/IF
1/100.
WB (1)
1/1000. Detects a band of approximately 58 kDa (predicted molecular weight: 56 kDa).
Notes
ICC/IF
1/100.
WB
1/1000. Detects a band of approximately 58 kDa (predicted molecular weight: 56 kDa).

Target

  • Function

    May play a role in the regulation of RNA biogenesis and/or mitosis. In vitro, dephosphorylates serine residues of skeletal muscle phosphorylase and histone H1.
  • Tissue specificity

    Ubiquitous.
  • Sequence similarities

    Belongs to the PPP phosphatase family. PP-5 (PP-T) subfamily.
    Contains 3 TPR repeats.
  • Cellular localization

    Nucleus. Cytoplasm. Predominantly nuclear. But also present in the cytoplasm.
  • Target information above from: UniProt accession P53041 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 5536 Human
    • Entrez Gene: 19060 Mouse
    • Entrez Gene: 65179 Rat
    • Omim: 600658 Human
    • SwissProt: P53041 Human
    • SwissProt: Q60676 Mouse
    • SwissProt: P53042 Rat
    • Unigene: 654604 Human
    • Unigene: 3294 Mouse
    • Unigene: 6107 Rat
    see all
  • Alternative names

    • FLJ36922 antibody
    • FLJ55954 antibody
    • OTTHUMP00000165834 antibody
    • PP-T antibody
    • PP5 antibody
    • PPP5 antibody
    • PPP5_HUMAN antibody
    • Ppp5c antibody
    • PPT antibody
    • Protein phosphatase 5, catalytic subunit antibody
    • Protein phosphatase T antibody
    • Serine/threonine protein phosphatase 5 antibody
    • Serine/threonine-protein phosphatase 5 antibody
    see all

Images

  • Western blot - Anti-PP-T antibody [12F7] (ab223367)
    Western blot - Anti-PP-T antibody [12F7] (ab223367)
    All lanes : Anti-PP-T antibody [12F7] (ab223367) at 1/1000 dilution

    Lane 1 : Wild-type HEK293T cell lysate
    Lane 2 : PPP5C knockout HEK293T cell lysate
    Lane 3 : A431 cell lysate
    Lane 4 : Jurkat cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) at 1/10000 dilution

    Predicted band size: 56 kDa
    Observed band size: 58 kDa why is the actual band size different from the predicted?



    Lanes 1-4: Merged signal (red and green). Green - ab223367 observed at 58 kDa. Red - loading control ab181602 observed at 36 kDa.

     ab223367 Anti-PP-T antibody [12F7] was shown to specifically react with PP-T in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266507 (knockout cell lysate ab258136) was used. Wild-type and PP-T knockout samples were subjected to SDS-PAGE. ab223367 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-PP-T antibody [12F7] (ab223367)
    Immunocytochemistry/ Immunofluorescence - Anti-PP-T antibody [12F7] (ab223367)

    4% formaldehyde-fixed HeLa (human epithelial cell line from cervix adnocarcinoma) cells stained for PP-T (green) using ab223367 at 1/100 dilution for 60 minutes at room temperature in ICC/IF, followed by Goat Anti-Mouse ATTO 488 at 1/100 dilution for 60 minutes at room temperature. Counterstain: DAPI (blue) nuclear stain at 1/5000 for 5 minutes room temperature.

    (A) DAPI nuclear stain (B) ab223367 (C) Merge.

  • Western blot - Anti-PP-T antibody [12F7] (ab223367)
    Western blot - Anti-PP-T antibody [12F7] (ab223367)
    All lanes : Anti-PP-T antibody [12F7] (ab223367) at 1/500 dilution

    Lane 1 : A431 (human epidermoid carcinoma cell line) cell lysate
    Lane 2 : HEK-293 (human epithelial cell line from embryonic kidney) cell lysate
    Lane 3 : Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Mouse IgG HRP at 1/200 dilution

    Developed using the ECL technique.

    Predicted band size: 56 kDa
    Observed band size: 58 kDa why is the actual band size different from the predicted?


    Exposure time: 6 minutes


    Blocking buffer: 5% skim milk.

    Primary and secondary antibodies were incubated for 1 hour at room temperature.

  • Immunocytochemistry/ Immunofluorescence - Anti-PP-T antibody [12F7] (ab223367)
    Immunocytochemistry/ Immunofluorescence - Anti-PP-T antibody [12F7] (ab223367)

    2% formaldehyde-fixed HEK-293 (human epithelial cell line from embryonic kidney) cells stained for PP-T (green) using ab223367 at 1/50 dilution in ICC/IF, followed by Goat Anti-Mouse Alexa Fluor 488 at 1/100 dilution for 60 minutes at RT. Counterstain: DAPI (blue) nuclear stain.

    (A) DAPI nuclear stain (B) ab223367 (C) Merge.

Protocols

  • Western blot protocols
  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (0)

Publishing research using ab223367? Please let us know so that we can cite the reference in this datasheet.

ab223367 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

Western blot abreview for Anti-PP-T antibody [12F7]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (mouse tissues)
Gel Running Conditions
Reduced Denaturing
Loading amount
50 µg
Specification
mouse tissues
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

Submitted Oct 19 2018

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