Overview

  • Product name

    Anti-Uroplakin III antibody [SFI-1]
    See all Uroplakin III primary antibodies
  • Description

    Mouse monoclonal [SFI-1] to Uroplakin III
  • Host species

    Mouse
  • Tested applications

    Suitable for: IHC-Fr, IHC-P, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Cow, Human, Common marmoset
  • Immunogen

    Synthetic peptide corresponding to an internal human Uroplakin III.

  • Positive control

    • Urothelial tissue, bladder.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.05% Proclin
    Constituents: 1% BSA, 0.58% Sodium chloride, 0.1% Tween, 0.363% Tris

    Containing antibody stabilizer
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Monoclonal
  • Clone number

    SFI-1
  • Isotype

    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab78196 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
IHC-Fr Use at an assay dependent concentration. PubMed: 21055807
IHC-P 1/20 - 1/30. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Incubate 30 minutes at room temperature.
WB Use at an assay dependent concentration. Predicted molecular weight: 32 kDa.

Target

  • Function

    Component of the asymmetric unit membrane (AUM); a highly specialized biomembrane elaborated by terminally differentiated urothelial cells. May play an important role in AUM-cytoskeleton interaction in terminally differentiated urothelial cells. It also contributes to the formation of urothelial glycocalyx which may play an important role in preventing bacterial adherence.
  • Tissue specificity

    Expressed in ureter.
  • Involvement in disease

    Defects in UPK3A are a cause of renal adysplasia (RADYS) [MIM:191830]; also known as renal agenesis or renal aplasia. Renal agenesis refers to the absence of one (unilateral) or both (bilateral) kidneys at birth. Bilateral renal agenesis belongs to a group of perinatally lethal renal diseases, including severe bilateral renal dysplasia, unilateral renal agenesis with contralateral dysplasia and severe obstructive uropathy.
  • Sequence similarities

    Belongs to the uroplakin-3 family.
  • Cellular localization

    Endoplasmic reticulum membrane. Heterodimer formation with UPK1B is a prerequisite to exit out of the endoplasmic reticulum (ER).
  • Information by UniProt
  • Database links

  • Alternative names

    • MGC119178 antibody
    • OTTHUMP00000028951 antibody
    • UP III antibody
    • UP3a antibody
    • UPIII antibody
    • UPIIIA antibody
    • UPK 3 antibody
    • UPK 3A antibody
    • UPK3 antibody
    • UPK3A antibody
    • UPK3A_HUMAN antibody
    • Uroplakin 3A antibody
    • Uroplakin III antibody
    • Uroplakin-3a antibody
    • Uroplakin3A antibody
    • UroplakinIII antibody
    see all

Images

  • ab78196 staining Uroplakin III in mouse bladder tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in citric acid. Samples were incubated with primary antibody (1/600 in TBS/BSA/azide) for 16 hours at 21°C. A Biotin-conjugated goat anti-mouse IgG polyclonal (1/200) was used as the secondary antibody.

    Positivity of only the lumenal cytoplasm of the Umbrella cells is clear.
    As this is a mouse primary antibody, endogenous IgG will also be visualised in the interstitial spaces (red arrowheads), in Plasma cells (black arrowheads) and also in blood vessels if the animal has not been perfused-washed/fixed.

    See Abreview

  • ab78196 staining Uroplakin III in cow ureter tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in citric acid. Samples were incubated with primary antibody (1/300 in TBS/BSA/azide) for 16 hours at 21°C. A Biotin-conjugated goat anti-mouse IgG polyclonal (1/200) was used as the secondary antibody.

    Image shows clear, strong labelling of the lumenal membranes of the Umbrella cells.

    See Abreview

  • IHC-P image of Uroplakin-III staining on Marmoset bladder sections using ab78196 (1:400). The sections were deparaffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10 mins at 21°C. ab78196 was diluetd 1:400 and incubted with the sections for 2 hours at 21°C. The secondary antibody used was Goat polyclonal to anti mouse IgG conjugated to biotin (1:200)

     

  • IHC-P image of Uroplakin-III staining on Rat bladder sections using ab78196 (1:400). The sections were deparaffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10 mins at 21°C. ab78196 was diluetd 1:1000 and incubted with the sections for 2 hours at 21°C. The secondary antibody used was Goat polyclonal to anti mouse IgG conjugated to biotin (1:250)

  • ab78196 at 1/20 dilution staining Uroplakin III in human bladder by Immunohistochemistry using formalin-fixed, paraffin-embedded tissue.

References

This product has been referenced in:

  • Fan G  et al. miR-33a hinders the differentiation of adipose mesenchymal stem cells towards urothelial cells in an inductive condition by targeting ß-catenin and TGFR. Mol Med Rep 17:2341-2348 (2018). Read more (PubMed: 29207162) »
  • Liu KM  et al. Ketamine-induced ulcerative cystitis and bladder apoptosis involve oxidative stress mediated by mitochondria and the endoplasmic reticulum. Am J Physiol Renal Physiol 309:F318-31 (2015). WB ; Rat . Read more (PubMed: 26109091) »
See all 3 Publications for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Bladder)
Permeabilization
Yes - Triton 0.1%
Specification
Bladder
Blocking step
CASBlock as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted May 12 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Sample
Goat Tissue sections (Ureter)
Specification
Ureter
Permeabilization
No
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Jun 02 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Sample
Cat Tissue sections (Ureter)
Specification
Ureter
Permeabilization
No
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted May 30 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Sample
Mouse Tissue sections (Bladder)
Specification
Bladder
Permeabilization
No
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Jan 14 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Sample
Cow Tissue sections (Ureter)
Specification
Ureter
Permeabilization
No
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Jan 14 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Sample
Rat Tissue sections (Bladder)
Specification
Bladder
Permeabilization
No
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Dec 06 2013

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Sample
Marmoset (common) Tissue sections (Bladder)
Specification
Bladder
Permeabilization
No
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Dec 05 2013

Answer

Thank you for your reply. It is possible that this band may be non-specific, or it may represent Uroplakin III in a different form. Forexample, if the protein is heavily glycosylated it may run higher than predicted. Alternatively, if isoform 2 of Uroplakin III were present in a heterodimer with Uroplakin 1b, the complex would be expected to run around 47 kDa. I would recommend running both positive and negative controls to determine whether the band is specific. Because Uroplakin III is specifically expressed in urothelial cells, you could use urinary bladder lysate as a positive control, and a negative control from an unrelated tissue (such as brain, liver, stomach, etc). Under optimized conditions, we would expect ab78196 to give a single band or a doublet (corresponding to the pro and mature forms) on urinary bladder, and no band in the negative control. Because Uroplakin III is expressed in the endoplasmic reticulum and membrane, it is important that your tissue is lysed in the presence of both ionic and non-ioninc detergents. You will be able to obtain the maximum specific signal by using a RIPA buffer for lysis. To avoid aggregation of membrane proteins, it can also often help to heat the samples to 70C for 15 minutes rather than boiling them. I hope this helps, please let me know if you need any additional assistance.

Read More

Question
Answer

Thank you for contacting us. According to the uniprot database (linked below), the predicted molecular weight of the Uroplakin III precursor, isoform 1, is 30.6 kDa. After the removal of the 18 amino acid signal peptide and the additional weight from several glycosylations, this protein will run around 32 kDa in a reduced and denatured western blot. It is possible that due to differences in buffer systems or differences in post-translational modifications between sample types, that this protein may appear at a slightly higher or lower molecular weight. There is also a smaller isoform which will run around 17 kDa. I hope this helps, please let me know if you need any additional information. http://www.uniprot.org/uniprot/O75631

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