Anti-Glypican 3 antibody (ab66596)

Rabbit polyclonal Glypican 3 antibody. Validated in WB, ELISA, IHC, ICC/IF and tested in Mouse, Human. Cited in 11 publication(s). Independently reviewed in 4 review(s).

Overview

  • Product name

  • Description

    Rabbit polyclonal to Glypican 3
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ELISA, IHC-P, ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat, Rabbit, Horse, Chicken, Guinea pig, Cow, Cat, Dog, Pig, Chimpanzee
  • Immunogen

    Synthetic peptide within Human Glypican 3 aa 300-349 (internal sequence). The exact sequence is proprietary. Numbering is based off mature sequence
    Sequence:

    FSTIHDSIQYVQKNAGKLTTTIGKLCAHSQQRQYRSAYYPEDLFIDKKVL


    Database link: P51654
    (Peptide available as ab111665)

  • Positive control

    • Placenta

Properties

Applications

Our Abpromise guarantee covers the use of ab66596 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
ELISA Use at an assay dependent concentration.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 2.5 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 66 kDa). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.

Target

  • Function

    Cell surface proteoglycan that bears heparan sulfate. Inhibits the dipeptidyl peptidase activity of DPP4. May be involved in the suppression/modulation of growth in the predominantly mesodermal tissues and organs. May play a role in the modulation of IGF2 interactions with its receptor and thereby modulate its function. May regulate growth and tumor predisposition.
  • Tissue specificity

    Highly expressed in lung, liver and kidney.
  • Involvement in disease

    Defects in GPC3 are the cause of Simpson-Golabi-Behmel syndrome type 1 (SGBS1) [MIM:312870]; also known as Simpson dysmorphia syndrome (SDYS). SGBS is a condition characterized by pre- and postnatal overgrowth (gigantism) with visceral and skeletal anomalies.
  • Sequence similarities

    Belongs to the glypican family.
  • Cellular localization

    Cell membrane and Secreted > extracellular space.
  • Information by UniProt
  • Database links

  • Alternative names

    • DGSX antibody
    • Glypican proteoglycan 3 antibody
    • Glypican-3 [Precursor] antibody
    • Gpc3 antibody
    • GPC3_HUMAN antibody
    • GTR2 2 antibody
    • GTR2-2 antibody
    • Heparan sulphate proteoglycan antibody
    • Intestinal protein OCI 5 antibody
    • Intestinal protein OCI-5 antibody
    • MXR7 antibody
    • OCI 5 antibody
    • OCI-5 antibody
    • OCI5 antibody
    • SDYS antibody
    • Secreted glypican-3 antibody
    • SGB antibody
    • SGBS antibody
    • SGBS1 antibody
    see all

Images

  • Anti-Glypican 3 antibody (ab66596) at 2.5 µg/ml (in 5% skim milk / PBS buffer) + Placenta lysate at 10 µg

    Secondary
    HRP conjugated anti-Rabbit IgG at 1/50000 dilution

    Predicted band size: 66 kDa
    Observed band size: 55 kDa
    why is the actual band size different from the predicted?



    Gel concentration: 12%
  • ICC/IF image of ab66596 stained Hek293 cells. The cells were 100% methanol fixed (5 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 (ab66596, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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.

  • IHC image of ab66596 staining in human placenta formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab66596, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References

This product has been referenced in:

  • Wei J  et al. Establishment and characterization of the GC-030-35 cell line derived from gastric hepatoid adenocarcinoma. Cancer Manag Res 11:1275-1287 (2019). Read more (PubMed: 30799954) »
  • Ni HM  et al. Receptor-Interacting Serine/Threonine-Protein Kinase 3 (RIPK3)-Mixed Lineage Kinase Domain-Like Protein (MLKL)-Mediated Necroptosis Contributes to Ischemia-Reperfusion Injury of Steatotic Livers. Am J Pathol N/A:N/A (2019). Read more (PubMed: 31026418) »
See all 14 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Cerebelum)
Antigen retrieval step
Heat mediated
Permeabilization
No
Specification
Cerebelum
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde

Miss. Virginia Gutiérrez

Verified customer

Submitted May 30 2016

Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (liver, hepatocellular carcinoma, Solt-Farber model)
Specification
liver, hepatocellular carcinoma, Solt-Farber model
Fixative
Acetone
Permeabilization
Yes - PBS-T
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Sep 13 2011

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Liver, hepatocellular carcinoma, Solt-Farber model)
Specification
Liver, hepatocellular carcinoma, Solt-Farber model
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: sodium citrate buffer pH 6.0
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Sep 13 2011

Application
Western blot
Sample
Mouse Tissue lysate - whole (Kidney)
Loading amount
30 µg
Specification
Kidney
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Dec 30 2010

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