Recombinant
RabMAb

Recombinant Anti-ALP antibody [EPR18663] (ab194297)

Overview

  • Product name
    Anti-ALP antibody [EPR18663]
    See all ALP primary antibodies
  • Description
    Rabbit monoclonal [EPR18663] to ALP
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ICC/IF, IP, IHC-P, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human ALP aa 800 to the C-terminus. The exact sequence is proprietary.
    Database link: Q9H0A0

  • Positive control
    • WB: Human fetal brain and fetal heart lysates; Mouse brain, rat heart and rat spleen lysates. IHC-P: Human colon, mouse stomach and rat colon tissues. ICC/IF: HeLa and NIH/3T3 cells. IP: HeLa cell lysate. FC: NIH/3T3 cell lysate
  • General notes

     

     

     This product was previously labelled as NAT10

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab194297 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 1/2000. Detects a band of approximately 116 kDa (predicted molecular weight: 116 kDa).
ICC/IF 1/2000.
IP 1/80.
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt 1/600.

Target

  • Function
    Has protein acetyltransferase activity in vitro. Can acetylate both histones and microtubules. Histone acetylation may regulate transcription and mitotic chromosome de-condensation. Activates telomerase activity by stimulating the transcription of TERT, and may also regulate telomerase function by affecting the balance of telomerase subunit assembly, disassembly, and localization. Acetylates alpha-tubulin, which may affect microtubule stability and cell division.
  • Sequence similarities
    Belongs to the UPF0202 family.
    Contains 1 N-acetyltransferase domain.
  • Cellular localization
    Nucleus > nucleolus. Nucleolar in interphase and redistributes to the perichromosomal layer and to the midbody during telophase.
  • Information by UniProt
  • Database links
  • Alternative names
    • ALP antibody
    • DKFZp434C116 antibody
    • FLJ10774 antibody
    • FLJ12179 antibody
    • FLJ23850 antibody
    • hALP antibody
    • KIAA1709 antibody
    • N acetyltransferase 10 antibody
    • N acetyltransferase 10 GCN5 related antibody
    • N acetyltransferase like antibody
    • N acetyltransferase like protein antibody
    • N-acetyltransferase 10 antibody
    • NAT10 antibody
    • NAT10_HUMAN antibody
    • NET43 antibody
    see all

Images

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling ALP with ab194297 at 1/2000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear staining on HeLa cell line.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:-
    -ve control 1: ab194297 at 1/2000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

     

  • All lanes : Anti-ALP antibody [EPR18663] (ab194297) at 1/2000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Rat heart lysate
    Lane 3 : Rat spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 116 kDa
    Observed band size: 116 kDa


    Exposure time: 30 seconds
  • Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling ALP with ab194297 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nucleus staining on rat colon tissue is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Flow cytometry analysis of NIH/3T3 (mouse embryo) cells labelling ALP (red) with purified ab194297 at dilution of 1/600. The secondary antibody used was Alexa Fluor® 488 goat-anti-rabbit IgG (1/2000). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Isotype control antibody used was Rabbit Monoclonal IgG (black). The blue line shows cells without incubation with primary antibody and secondary antibody.

  • All lanes : Anti-ALP antibody [EPR18663] (ab194297) at 1/2000 dilution

    Lane 1 : Human fetal brain lysate
    Lane 2 : Human fetal heart lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 116 kDa
    Observed band size: 116 kDa


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling ALP with ab194297 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nucleus staining on Human colon tissue is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embyro fibroblast cells) cells labeling ALP with ab194297 at 1/2000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear staining on NIH/3T3 cell line.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:-
    -ve control 1: ab194297 at 1/2000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

     

  • Immunohistochemical analysis of paraffin-embedded Mouse stomach tissue labeling ALP with ab194297 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nucleus staining on mouse stomach tissue is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • ALP was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) cell lysate with ab194297 at 1/80 dilution.

    Lane 1: HeLa cell lysate 10ug (Input).

    Lane 2: ab194297 IP in HeLa cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab194297 in HeLa cell lysate.

    Western blot was performed from the immunoprecipitate using ab194297 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 seconds.

References

ab194297 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (PC12, a cell line derived from a pheochromocytoma)
Permeabilization
Yes - 0.1% TritonX-100
Specification
PC12, a cell line derived from a pheochromocytoma
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde

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Submitted Sep 18 2017

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