Overview

  • Product name
  • Description
    Rabbit polyclonal to PP-X
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, WB, ICCmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human
    Predicted to work with: Rabbit
  • Immunogen

    Synthetic peptide corresponding to Human PP-X aa 294-307 conjugated to keyhole limpet haemocyanin.
    Sequence:

    RGIPSKKPVADYFL

  • Positive control
    • Recombinant Human PP-X protein (ab114692) can be used as a positive control in WB. Bovine testes homogenate.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.08% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab16475 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 1/300.
WB 1/500 - 1/1000. Predicted molecular weight: 34 kDa.
ICC 1/500 - 1/1000.

Target

  • Function
    Protein phosphatase that is involved in many processes such as microtubule organization at centrosomes, maturation of spliceosomal snRNPs, apoptosis, DNA repair, tumor necrosis factor (TNF)-alpha signaling, activation of c-Jun N-terminal kinase MAPK8, regulation of histone acetylation, DNA damage checkpoint signaling, NF-kappa-B activation and cell migration. The PPP4C-PPP4R1 PP4 complex may play a role in dephosphorylation and regulation of HDAC3. The PPP4C-PPP4R2-PPP4R3A PP4 complex specifically dephosphorylates H2AFX phosphorylated on Ser-140 (gamma-H2AFX) generated during DNA replication and required for DNA double strand break repair. Dephosphorylates NDEL1 at CDK1 phosphorylation sites and negatively regulates CDK1 activity in interphase (By similarity). In response to DNA damage, catalyzes RPA2 dephosphorylation, an essential step for DNA repair since it allows the efficient RPA2-mediated recruitment of RAD51 to chromatin.
  • Sequence similarities
    Belongs to the PPP phosphatase family. PP-4 (PP-X) subfamily.
  • Cellular localization
    Cytoplasm. Nucleus. Cytoplasm > cytoskeleton > centrosome.
  • Information by UniProt
  • Database links
  • Alternative names
    • PP X antibody
    • PP-X antibody
    • Pp4 antibody
    • PP4C antibody
    • PP4C_HUMAN antibody
    • PPH3 antibody
    • PPP4 antibody
    • ppp4c antibody
    • PPX antibody
    • protein phosphatase 4 (formerly X), catalytic subunit antibody
    • Protein phosphatase 4 catalytic subunit antibody
    • Protein phosphatase X antibody
    • protein phosphatase X, catalytic subunit antibody
    • Serine/threonine protein phosphatase 4 catalytic subunit antibody
    • Serine/threonine-protein phosphatase 4 catalytic subunit antibody
    see all

Images

  • All lanes : Anti-PP-X antibody (ab16475) at 1/10000 dilution (in PBS + 5% milk +0.1% Tween 20 for 1 hour at 25°C)

    Lane 1 : HeLa whole cell lysate - mock transfected
    Lane 2 : HeLa whole cell lysate - transfected with human siRNA against PP-X

    Lysates/proteins at 100000 cells per lane.

    Secondary
    All lanes : An HRP-conjugated Goat anti-rabbit IgG polyclonal at 1/50000 dilution

    Performed under non-reducing conditions.

    Predicted band size: 34 kDa
    Observed band size: 34 kDa
    Additional bands at: 50 kDa. We are unsure as to the identity of these extra bands.



    Blocking Step: 5% Milk for 1 hour at 25°C

    See Abreview

References

This product has been referenced in:
See 1 Publication for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Application
Western blot
Loading amount
10000 cells
Gel Running Conditions
Reduced Denaturing (4-12% gradient)
Sample
Human Cell lysate - whole cell (Huh7 liver)
Specification
Huh7 liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Mar 31 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Hela cells)
Loading amount
100000 cells
Specification
Hela cells
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (Invitrogen NuPage 4-12% Bis-Tris)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Sep 22 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Hela)
Specification
Hela
Fixative
Methanol
Permeabilization
No
Blocking step
Fish scale gelatin as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.2% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jul 15 2010

Question

I have a customer who purchased an anti-PPPC4 antibody from you, via Novus. I believe that the Novus PO# for this antibody is #BST031005R. At any rate, the customer claims that he did not obtain a signal from this antibody, although he ran a loading control (which worked). He has forwarded his protocol and images. Could you please tell me how you would like to handle this one? Thank you! ---- 1. Briefly describe the problem you are experiencing: There’s no signal, and I have tried several times. GAPDH can be detected with the same method and reagents and a different antibdoy. Detection species: Rat cerebrum; Many papers show target protein exists in this sample. http://bioinfo1.weizmann.ac.il/cgi-bin/genecards/carddisp?PPP4C&search=PP4c&suff=txt Have you used this antibody successfully in the past?: No____ Using first time ______ 2. How is this antibody stored? -20?C 3. Please describe how your sample was prepared, including species: my samples are cell extract, sample buffer formulation :RIPA, phosphotase inhibitor, protease inhibitor, PMSF 4. What type of gel was used?: 10%, 12%,15% all have tried 5. Please describe transfer conditions: 25mM tris base 0.2M glycine,20% methanol(PH=8.5) 6. Please describe blocking conditions:5% milk 7. Primary antibody: dilution 1:500, 1:200 have tried dilution buffer:TBST with 5%BSA incubation time and temperature: 4? overnight 8. Secondary antibody: dilution 1:1000 dilution buffer:5% milk incubation time and temperature: 1 h rt Was it tested using another primary antibody?: yes 9. What detection system did you use?: ECL Phototope®-HRP detection kit (Cell signal Technology) 10. Did you use a positive control and what where the results? No, because I don’t have one, but there are many papers showing the target protein exists in this sample. http://bioinfo1.weizmann.ac.il/cgi-bin/genecards/carddisp?PPP4C&search=PP4c&suff=txt 11. Did you use a negative control and what where the results? no 12. Results: Did you see any bands? No Where there any unexpected bands? Please describe any addition observations: _Please take a look at the picture of western blotting.

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Answer

I think there are two potential areas which the customer should consider looking into: the protocol and the samples. I am sure the customer knows very well his area of research and trust that many papers show PPP4C where he expects it, unfortunately, not all antibodies are the same and some detect even small amounts of protein while others are not so ideal, so I would recommend to run our recommended positive control of bovine testes homogenate. As you know our policy is that we offer a replacement or refund if the antibody does not perform as stated on the datasheet (and provided the antibody was purchased within 90 days prior to the complaint) so I will be certainly be able to offer this guarantee if the antibody does not work on this positive control. Please also note PP4C is localised both in the cytoplasm and the nucleus, maybe in the customer's samples the protein is concentrated in the nucleus (a nuclear extraction may therefore be necessary). The second area to investigate is the protocol. It looks very good and I think the customer has near-optimised it, however we have numerous occasions where the blocking agent is responsible for no signal, (too much of a good thing) and actually binds the membrane so well that the antibody can't bind to it too, and I would'nt recommend mixing BSA and milk in the same experiment. The customer should try several blocking experiments: BSA 5% (1hr) and no blocking in the dilution buffer, and milk 5% (1hr) and no blocking in the dilution buffer. If the customer still experiences problems with the bovine testes samples please do not hesitate to contact me again and we will arrange a replacement if the antibody was bought in the last 90days.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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