Overview

  • Product name

    Anti-GOLPH3/MIDAS antibody
    See all GOLPH3/MIDAS primary antibodies
  • Description

    Rabbit polyclonal to GOLPH3/MIDAS
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IFmore details
  • Species reactivity

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

    Synthetic peptide corresponding to Human GOLPH3/MIDAS aa 1-100 (N terminal) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab108475)

  • Positive control

    • This antibody gave a positive signal in Human, Mouse and Rat Lung Tissue Lysates, and in HeLa and A549 Whole Cell Lysates.
  • General notes

     This product was previously labelled as GOLPH3

     

Properties

Applications

Our Abpromise guarantee covers the use of ab98023 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 a concentration of 1 µg/ml. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa).
ICC/IF Use a concentration of 5 µg/ml.

Target

  • Function

    Involved in modulation of mTOR signaling. Involved in the regulation of mitochondrial lipids, leading to increase of mitochondrial mass. Potential oncogene.
  • Tissue specificity

    Detected in muscle fibers of patients with mitochondrial diseases; not detected in normal muscle fibers.
  • Sequence similarities

    Belongs to the GOLPH3/VPS74 family.
  • Post-translational
    modifications

    Phosphorylated.
  • Cellular localization

    Cell membrane. Endosome. Cytoplasm. Golgi apparatus > Golgi stack membrane. Mitochondrion intermembrane space.
  • Information by UniProt
  • Database links

  • Alternative names

    • Coat protein antibody
    • Coat protein GPP34 antibody
    • FLJ90675 antibody
    • Golgi associated protein antibody
    • Golgi peripheral membrane protein 1, 34 kDa antibody
    • Golgi phosphoprotein 3 (coat protein) antibody
    • Golgi phosphoprotein 3 antibody
    • Golgi protein antibody
    • GOLP3_HUMAN antibody
    • Golph3 antibody
    • GOPP1 antibody
    • GPP34 antibody
    • MIDAS antibody
    • Mitochondrial DNA absence factor antibody
    see all

Images

  • All lanes : Anti-GOLPH3/MIDAS antibody (ab98023) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Lung (Human) Tissue Lysate
    Lane 3 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate
    Lane 4 : Mouse lung normal tissue lysate - total protein (ab29297)
    Lane 5 : Lung (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 34 kDa
    Observed band size: 34 kDa
    Additional bands at: 32 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 30 seconds


    The predicted molecular weight of GOLPH3/MIDAS is 34-kDa. We are unsure why we are seeing an additional band in HeLa and A549 whole cell lysates. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

  • ab98023 staining GOLPH3/MIDAS in HeLa cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X100. Samples were incubated with ab98023 (1/200: in PBS) for 1 hours at 22°C. An Alexa Fluor®488-conjugated goat polyclonal to rabbit IgG was used at dilution at 1/200 as secondary antibody. The nuclei are counterstained with DAPI.

    See Abreview

  • ICC/IF image of ab98023 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab98023, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2 and MCF7 cells at 5µg/ml.

References

This product has been referenced in:

  • Arriagada C  et al. The knocking down of the oncoprotein Golgi phosphoprotein 3 in T98G cells of glioblastoma multiforme disrupts cell migration by affecting focal adhesion dynamics in a focal adhesion kinase-dependent manner. PLoS One 14:e0212321 (2019). Read more (PubMed: 30779783) »
  • Liu H  et al. Golgi phosphoprotein 3 (GOLPH3) promotes hepatocellular carcinoma progression by activating mTOR signaling pathway. BMC Cancer 18:661 (2018). Read more (PubMed: 29914442) »
See all 4 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

Filter by Ratings

1-2 of 2 Abreviews

Application
Western blot
Sample
Human Cell lysate - whole cell (U2OS)
Gel Running Conditions
Reduced Denaturing (4-20% tris-glycine)
Loading amount
30 cells
Specification
U2OS
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Jun 17 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - 0.5% Triton X100
Specification
HeLa
Fixative
Paraformaldehyde

Dr. Kirk Mcmanus

Verified customer

Submitted May 25 2011

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up