Overview

  • Product name

  • Description

    Rabbit polyclonal to PAR-3/PARD3
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-FrFl, ICC/IF, IHC-Pmore details
  • Species reactivity

    Reacts with: Rat, Human
    Predicted to work with: Mouse, Sheep, Chimpanzee
  • Immunogen

    Synthetic peptide corresponding to Human PAR-3/PARD3 aa 1300 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab73313, ab73314)

  • Positive control

    • This antibody gave a positive signal in the following Lysates: HeLa Whole Cell, Rat Thymus Tissue, Rat Brain Tissue, and SW480 Whole Cell This antibody gave a positive result in IHC in the following FFPE tissue: Human lung adenocarcinoma.
  • General notes

    Previously labelled as PARD3

Properties

Applications

Our Abpromise guarantee covers the use of ab64646 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 150 kDa (predicted molecular weight: 150 kDa).
IHC-FrFl Use at an assay dependent concentration.
ICC/IF Use a concentration of 1 - 5 µg/ml.
IHC-P Use a concentration of 0.5 µg/ml.

Target

  • Function

    Adapter protein involved in asymmetrical cell division and cell polarization processes. Seems to play a central role in the formation of epithelial tight junctions. Targets the phosphatase PTEN to cell junctions (By similarity). Association with PARD6B may prevent the interaction of PARD3 with F11R/JAM1, thereby preventing tight junction assembly. The PARD6-PARD3 complex links GTP-bound Rho small GTPases to atypical protein kinase C proteins. Required for establishment of neuronal polarity and normal axon formation in cultured hippocampal neurons.
  • Tissue specificity

    Widely expressed.
  • Sequence similarities

    Belongs to the PAR3 family.
    Contains 3 PDZ (DHR) domains.
  • Domain

    Contains a conserved N-terminal oligomerization domain (NTD) that is involved in oligomerization and is essential for proper subapical membrane localization.
    The second PDZ domain mediates interaction with membranes containing phosphoinositol lipids.
  • Post-translational
    modifications

    Phosphorylated by PRKCZ. EGF-induced Tyr-1127 phosphorylation mediates dissociation from LIMK2.
    Phosphorylation by STK6/AURKA at Ser-962 is required for the normal establishment of neuronal polarity.
  • Cellular localization

    Endomembrane system. Cell junction. Cell junction > tight junction. Cell membrane. Cytoplasm > cell cortex. Cytoplasm > cytoskeleton. Localized along the cell-cell contact region. Colocalizes with PARD6A and PRKCI at epithelial tight junctions. Colocalizes with the cortical actin that overlays the meiotic spindle during metaphase I and metaphase II (By similarity). Presence of KRIT1, CDH5 and RAP1B is required for its localization to the cell junction.
  • Information by UniProt
  • Database links

  • Alternative names

    • agouti signaling protein antibody
    • ASIP antibody
    • Atypical PKC isotype specific interacting protein antibody
    • Atypical PKC isotype-specific-interacting protein antibody
    • Baz antibody
    • bazooka antibody
    • Coagulation factor II receptor-like 2 antibody
    • CTCL tumor antigen se2-5 antibody
    • F2rl2 antibody
    • PAR-3 antibody
    • par-3 family cell polarity regulator alpha antibody
    • par-3 family cell polarity regulator antibody
    • PAR3 alpha antibody
    • par3 antibody
    • PAR3-alpha antibody
    • PARD-3 antibody
    • Pard3 antibody
    • PARD3_HUMAN antibody
    • PARD3A antibody
    • Partitioning defective 3 homolog antibody
    • Partitioning-defective 3 homolog antibody
    • Partitioning-defective protein 3, C. elegans, homolog of antibody
    • SE2-5L16 antibody
    • SE2-5LT1 antibody
    • SE2-5T2 antibody
    • Thrombin receptor-like 2 antibody
    see all

Images

  • IHC image of PAR-3/PARD3 staining in Human lung adenocarcinoma 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 ab64646, 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.

  • ab64646 stained A549 cells. The cells were 100% methanol fixed for 5 minutes at-20°C* and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab64646 at 5µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • All lanes : Anti-PAR-3/PARD3 antibody (ab64646) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Thymus (Rat) Tissue Lysate
    Lane 3 : Brain (Rat) Tissue Lysate
    Lane 4 : SW480 (Human colon adenocarcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 150 kDa
    Observed band size: 150 kDa
    Additional bands at: 170 kDa, 45 kDa, 53 kDa, 60 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 1 minute

References

This product has been referenced in:

  • Stypulkowski E  et al. The depalmitoylase APT1 directs the asymmetric partitioning of Notch and Wnt signaling during cell division. Sci Signal 11:N/A (2018). Read more (PubMed: 29295957) »
  • Song T  et al. Loss of Par3 promotes lung adenocarcinoma metastasis through 14-3-3? protein. Oncotarget 7:64260-64273 (2016). Read more (PubMed: 27588399) »
See all 4 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 (duodenum)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: ph9 steamer
Permeabilization
No
Specification
duodenum
Blocking step
BSA as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Fixative
Formaldehyde

Miss. Anne Sailer

Verified customer

Submitted Aug 04 2015

Answer

Thanks for your enquiry! I will enquire with our supplying lab to determine if ab64840 may also cross react with zebrafish PARD3 and will forward you any information as soon as possible. As we discussed, please reply to this email to let me know if you are interested in participating in our testing discount program.

Read More
Application
Immunohistochemistry free floating
Sample
Rat Tissue sections (Brain)
Specification
Brain

Dr. Sophie Pezet

Verified customer

Submitted Jul 29 2011

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
No
Specification
HeLa
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 27°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Sep 09 2010

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