Overview

  • Product name

  • Description

    Rabbit polyclonal to TAZ
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Human
    Predicted to work with: Rat, Rabbit, Guinea pig, Cow, Dog
  • Immunogen

    Synthetic peptide within Human TAZ aa 125-121 (N terminal). The exact sequence is proprietary.
    Sequence:

    PRLAGGAQHVRSHSSPASLQLGTGAGAAGSPAQQHAHLRQQSYDVTDELP


    Database link: Q9GZV5

  • Positive control

    • HeLa whole cell lysate (ab150035) This antibody gave a positive result in IHC in the following FFPE tissue: Human normal lung.

Properties

Applications

Our Abpromise guarantee covers the use of ab84927 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-P Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 44 kDa. Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
ICC/IF Use at an assay dependent concentration.

Target

  • Function

    Transcriptional coactivator which acts as a downstream regulatory target in the Hippo signaling pathway that plays a pivotal role in organ size control and tumor suppression by restricting proliferation and promoting apoptosis. The core of this pathway is composed of a kinase cascade wherein MST1/MST2, in complex with its regulatory protein SAV1, phosphorylates and activates LATS1/2 in complex with its regulatory protein MOB1, which in turn phosphorylates and inactivates YAP1 oncoprotein and WWTR1/TAZ. WWTR1 enhances PAX8 and NKX2-1/TTF1-dependent gene activation. Regulates the nuclear accumulation of SMADS and has a key role in coupling them to the transcriptional machinery such as the mediator complex. Regulates embryonic stem-cell self-renewal, promotes cell proliferation and epithelial-mesenchymal transition.
  • Tissue specificity

    Highly expressed in kidney, heart, placenta and lung. Expressed in the thyroid tissue.
  • Sequence similarities

    Contains 1 WW domain.
  • Domain

    The PDZ-binding motif is essential for stimulated gene transcription. It localizes the protein into both punctate nuclear foci and plasma membrane-associated complexes.
    Binds to transcription factors via its WW domain.
  • Post-translational
    modifications

    Phosphorylated by LATS2 and STK3/MST2. Phosphorylation by LATS2 results in creation of 14-3-3 binding sites, retention in the cytoplasm, and functional inactivation. Phosphorylation results in the inhibition of transcriptional coactivation through YWHAZ-mediated nuclear export.
  • Cellular localization

    Nucleus. Cytoplasm. Concentrates along specific portions of the plasma membrane, and accumulates in punctate nuclear bodies. When phosphorylated, is retained in cytoplasm by YWHAZ. Can be retained in the nucleus by MED15.
  • Information by UniProt
  • Database links

  • Alternative names

    • DKFZP586I1419 antibody
    • FLJ27004 antibody
    • FLJ45718 antibody
    • OTTHUMP00000215994 antibody
    • OTTHUMP00000215995 antibody
    • OTTHUMP00000215996 antibody
    • OTTHUMP00000216001 antibody
    • TAZ antibody
    • Transcriptional co activator with PDZ binding motif antibody
    • Transcriptional coactivator with PDZ binding motif antibody
    • Transcriptional coactivator with PDZ-binding motif antibody
    • WW domain containing transcription regulator 1 antibody
    • WW domain containing transcription regulator protein 1 antibody
    • WW domain-containing transcription regulator protein 1 antibody
    • WWTR 1 antibody
    • WWTR1 antibody
    • WWTR1_HUMAN antibody
    see all

Images

  • All lanes : Anti-TAZ antibody (ab84927) at 1 µg/ml

    Lane 1 : HEK293 over-expression
    Lane 2 : Empty vector
    Lane 3 : Human heart - overexpression

    Predicted band size: 44 kDa



    Blocking buffer used was 3% milk 0.05% TBST

  • IHC image of TAZ staining in Human normal lung 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 ab84927, 1µg/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.

  • Immunocytochemical analysis of N1E-115 murine neuroblastoma cell line, labeling TAZ with ab84927. Cells were fixed in paraformaldehyde and permeabilized in 0.1% Triton X100. Blocking was performed with 4% BSA for 30 minutes at 21°C. Incubation with ab84927 diluted 1/100 in 4% BSA for 45 mintues at 21°C.  

    See Abreview

  • Anti-TAZ antibody (ab84927) at 1/1000 dilution + MDA-MB-231 breast cancer cells at 50000 cells with 2% milk + 1% BSA

    Performed under reducing conditions.

    Predicted band size: 44 kDa

    See Abreview

References

This product has been referenced in:

  • Kingston NM  et al. Immunofluorescence Microscopy to Study Endogenous TAZ in Mammalian Cells. Methods Mol Biol 1893:107-113 (2019). Read more (PubMed: 30565129) »
  • Zhao W  et al. Limonin attenuates the stemness of cervical carcinoma cells by promoting YAP nuclear-cytoplasmic translocation. Food Chem Toxicol 125:621-628 (2019). Read more (PubMed: 30738134) »
See all 14 Publications for this product

Customer reviews and Q&As

1-10 of 12 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Trachea)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate pH 6
Specification
Trachea
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Aug 16 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HEK293T)
Permeabilization
Yes - 0.1% Triton X-100
Specification
HEK293T
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jul 17 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (Vascular Smooth Muscle Cell)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
Vascular Smooth Muscle Cell
Blocking step
Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted May 16 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Dog Cell (Madine-Darby Canine Kidney cells)
Permeabilization
Yes - NP40
Specification
Madine-Darby Canine Kidney cells
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 15 2016

Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing (10% SDS PAGE)
Sample
Mouse Tissue lysate - whole (Liver)
Specification
Liver
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jul 15 2014

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (N1E-115 murine neuroblastoma)
Specification
N1E-115 murine neuroblastoma
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton X100
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Feb 13 2013

Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (MDA-MB-231 breast cancer cells)
Loading amount
50000 cells
Specification
MDA-MB-231 breast cancer cells
Gel Running Conditions
Reduced Denaturing (10% PAGE)
Blocking step
2% milk + 1% BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Oct 05 2012

Answer

Thank you for your response.



This is to let you know that I have confirmed and agreed on placing a new order for your customer - for one vial ofab119373 ((exchange for the original ab84927)) as a free of charge replacement. I hope the second vial will work as it is expected, and please do let me know how you are getting on with this product.





Dear Colleague,



Please add 1 vial ofab119373 (as an exchange for ab84927) to the next shipment for Sapphire.

Original CCE ID: 3437495

Original Order: 983749

Problem is: No band in Western blot -Striping and reprobing membranes for β-actin clearly shows bands at the right size

Many thanks.

Read More

Question

LOT NUMBER GR7518 ORDER NUMBER PO-12393 DESCRIPTION OF THE PROBLEM No bands (not even non-specific) SAMPLE Human melanoma cell line lysates, MCF7 lysate, HCT116 lysate, human ovarian cancer cell line lysates and HeLa cell lysate PRIMARY ANTIBODY Polyclonal rabbit anti-Taz antibody (ab84927) used in 1µg/mL in 5% milk/PBS. 9mL made and put on membrane overnight on shaker at 4˚C. Washed off primary antibody in 3x 5 minute washes in PBST. DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED No lysates as negative control and HeLa lysates as positive control used as specified by the data sheet. ANTIBODY STORAGE CONDITIONS Aliquoted into -20˚C SAMPLE PREPARATION Cell lines lysed in RIPA buffer with phosphatase and protease inhibitors. Boiled samples in denaturing loading buffer AMOUNT OF PROTEIN LOADED Between 20-60µg ELECTROPHORESIS/GEL CONDITIONS Reducing SDS-PAGE, gradient gel of 6-15% TRANSFER AND BLOCKING CONDITIONS Wet transfer to PVDF membrane in transfer buffer containing glycine, tris and methanol for 1 hour at 100V. Blocking in 5% milk/PBS for 30 minutes. SECONDARY ANTIBODY Goat anti-rabbit-HRP from Santa Cruz diluted to 1:10 000 in 5% milk/PBS. Secondary antibody incubation for 1 hour on shaker at room temperature. Washed off secondary antibody in 3x5 minute washes in PBST. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? I’ve changed the amount of lysate loaded from 20, 40 and 60µg-but still no bands. Changed concentration of the primary antibody to 2µg/mL and also changed the dilution of the secondary antibody to 1:5000, but still no bands. Also tried ECL advanced (Amersham) but extremely faint bands show up at around 50-55kD, but no other bands detected. ADDITIONAL NOTES Striping and reprobing membranes for β-actin clearly shows bands at the right size, but with the Taz antibody, there are absolutely no bands, not even non-specific bands. Abcam invoice number: 1297530F

Read More
Answer

Thank you for your enquiry regarding ab84927 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody. The protocol looks absolutely fine to me and since after reprobingthe membranes there was a clear signal withβ-actin it implies that the protein transfer was fine. It may well be that the antibody has been damaged accidentally during transfer. I could offer your troubled customer either a new vial as a free of charge replacement or a credit note which can be used in the future. I look forward to hearing from you soon.

Read More

Question
Answer

Gracias por contactarnos. Basándose en la secuencia del péptido usado como inmunógeno, este anticuerpo comparte 100% de homología con la proteína TAZ y tan solo un 65% de homología con YAP1 (isoformas 1-3). Si lo necesitas puedo preguntar por los otros tres anticuerpos anti TAZ que tenemos en el catálogo testados en WB en especie humana, pero me atrevería a decir que el porcentaje de homología entre el péptido utilizado para producir el anticuerpo y la proteína YAP debe rondar el porcentaje mencionado, alrededor del 60%. Espero haberos ayudado, si necesitáis cualquier otra información, no dudéis en contactarnos de nuevo.

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1-10 of 12 Abreviews or Q&A

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