Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR13439] to GOLPH4/GPP130
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-GOLPH4/GPP130 antibody [EPR13439]
    See all GOLPH4/GPP130 primary antibodies

  • Description

    Rabbit monoclonal [EPR13439] to GOLPH4/GPP130

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-Pmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide within Human GOLPH4/GPP130 aa 50-150. The exact sequence is proprietary.
    Database link: O00461

  • Positive control

    • WB: HeLa, HEK-293T and HepG2 whole cell lysates. IHC-P: Human renal adenocarcinoma tissue.

  • General notes

     

     

     This product was previously labelled as GOLPH4

     

    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.

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

    pH: 7.2
    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

    EPR13439

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab197595 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 1/5000. Detects a band of approximately 130 kDa (predicted molecular weight: 82 kDa).
IHC-P 1/500 - 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • Function

    Plays a role in endosome to Golgi protein trafficking; mediates protein transport along the late endosome-bypass pathway from the early endosome to the Golgi.

  • Sequence similarities

    Belongs to the GOLIM4 family.

  • Post-translational
    modifications

    Phosphorylated probably by c-AMP-dependent kinases in its lumenal part.
    O-glycosylated; modified by sialic acid residues.
    N-glycosylated; N-glycans are probably of the complex type and modified by sialic acid residues.

  • Cellular localization

    Golgi apparatus > Golgi stack membrane. Endosome membrane. Localizes to cis and medial Golgi cisternae. Probably cycles between early Golgi and distal compartments like endosome.
  • Information by UniProt

  • Database links

    • Alternative names

      • 130 kDa golgi localized phosphoprotein antibody
      • cis antibody
      • cis Golgi localized calcium binding protein antibody
      • GIMPc antibody
      • Golgi integral membrane protein 4 antibody
      • Golgi integral membrane protein antibody
      • Golgi phosphoprotein 4 antibody
      • Golgi phosphoprotein of 130 kDa antibody
      • Golgi-localized phosphoprotein of 130 kDa antibody
      • GOLI4_HUMAN antibody
      • golim4 antibody
      • GOLPH 4 antibody
      • GPP 130 antibody
      • GPP130 antibody
      • P 138 antibody
      • P138 antibody
      • type II Golgi membrane protein antibody
      see all

    Images

    • Western blot - Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595)
      Western blot - Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595)
      All lanes : Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595) at 1/1000 dilution

      Lane 1 : Wild-type HEK293T cell lysate
      Lane 2 : GOLIM4 knockout HEK293T cell lysate
      Lane 3 : HeLa cell lysate
      Lane 4 : MOLT-4 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: 82 kDa
      Observed band size: 130 kDa
      why is the actual band size different from the predicted?



      Lanes 1-4: Merged signal (red and green). Green - ab197595 observed at 130 kDa. Red - loading control ab8245 observed at 36 kDa.

      ab197595 Anti-GOLPH4/GPP130 antibody [EPR13439] was shown to specifically react with GOLPH4/GPP130 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266123 (knockout cell lysate ab257455) was used. Wild-type and GOLPH4/GPP130 knockout samples were subjected to SDS-PAGE. ab197595 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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® 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.

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595)

      Immunohistochemical analysis of paraffin-embedded Human renal adenocarcinoma tissue labeling GOLPH4/GPP130 with ab197595 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasm staining on Human renal adenocarcinoma tissue is observed. Counter stained with Hematoxylin.

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

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Western blot - Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595)
      Western blot - Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595)
      Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595) at 1/2500 dilution + HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 20 µg

      Secondary
      Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

      Predicted band size: 82 kDa
      Observed band size: 130 kDa why is the actual band size different from the predicted?


      Exposure time: 1 minute


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The observed MW is consistent with what has been described in the literature PMID:20130081 and PMID:9201717

    • Western blot - Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595)
      Western blot - Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595)
      All lanes : Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595) at 1/5000 dilution

      Lane 1 : HepG2(Human liver hepatocellular carcinoma) whole cell lysate
      Lane 2 : 293(Human embryonic kidney) whole cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

      Predicted band size: 82 kDa
      Observed band size: 130 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The observed MW is consistent with what has been described in the literature (PMID:)20130081 and 9201717

    References (1)

    Publishing research using ab197595? Please let us know so that we can cite the reference in this datasheet.

    ab197595 has been referenced in 1 publication.

    • Bonsergent E & Lavieu G Content release of extracellular vesicles in a cell-free extract. FEBS Lett 593:1983-1992 (2019). PubMed: 31175663

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