リコンビナント
RabMAb

Anti-Pax2 抗体 [EP3251] (ab79389)

製品の概要

  • 製品名
    Anti-Pax2 antibody [EP3251]
    Pax2 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EP3251] to Pax2
  • 由来種
    Rabbit
  • アプリケーション
    適用あり: WB, Flow Cyt, IHC-P, ICC/IFmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide within Human Pax2 aa 1-100. The exact sequence is proprietary.
    Database link: Q02962
    (Peptide available as ab188213)

  • ポジティブ・コントロール
    • WB: Human fetal kidney tissue lysate IHC-P: Human fetal kidney, human normal kidney, human renal cell carcinoma, mouse liver and rat cerebral cortex tissues. ICC/IF: HEK293 cells. Flow Cyt: HEK293 and K562 cells.
  • 特記事項

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab79389 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
WB 1/1000 - 1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 45 kDa).Can be blocked with Pax2 peptide (ab188213).
Flow Cyt 1/50.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P 1/500 - 1/2500.

See IHC antigen retrieval protocols.

ICC/IF 1/50.

ターゲット情報

  • 関連性
    Pax2 is a transcription factor critically required during the development of the nervous and excretory systems, including the midbrain, hindbrain, spinal cord, eye, ear and urogenital tract. Like other products of the Pax gene family, Pax2 encodes a conserved 128 amino acid paired box DNA-binding domain in the N-terminal portion of the molecule. Function: Probable transcription factor that may have a role in kidney cell differentiation. Has a critical role in the development of the urogenital tract, the eyes, and the CNS. Tissue specificity: Expressed in primitive cells of the kidney, ureter, eye, ear and central nervous system.
  • 細胞内局在
    Nuclear
  • 参照データベース
  • 別名
    • FSGS7 antibody
    • Paired box 2 antibody
    • Paired box gene 2 antibody
    • paired box homeotic gene 2 antibody
    • paired box protein 2 antibody
    • Paired box protein Pax 2 antibody
    • Paired box protein Pax-2 antibody
    • Paired box protein Pax2 antibody
    • PAPRS antibody
    • Pax 2 antibody
    see all

画像

  • Pax2 is expressed in melanocytes of benign nevi (A) and melanoma cells of patients with malignant melanoma (B).

    Cells were grown on coverslips and fixed with 4% paraformaldehyde/PBS. After washing the cells with PBS, cells were permeabilized and blocked with 0.1% Triton X-100/PBS containing 5% BSA. Pax2 (green) was examined by immunofluorescent analysis using ab79389 at 1/100 dilution (incubated for 1 hour at room temperature). Following 3 times washing, bound antibodies were deteced by Alexa 488 conjugated goat anti-mouse or Cy3 conjugated goat anti-mouse secondary antibodies.

    Following PBS-washing nuclei were stained with 4′6-diamidino-2-phenylindole (DAPI, Blue) and cells were mounted in Fluoromount-G™ and examined by fluorescence microscopy.

    White arrows in the higher magnified insets indicate PAX2 expression in nucleoli of melanocytes of benign nevi (A), yellow arrows in the higher magnified insets specify PAX2 expression in nucleoli of melanoma cells (B).

  • Immunohistochemical analysis of Pax2 expression in tissue sections of benign nevi and malignant melanoma using ab79389 at 1/100 dilution.

    All specimens were fixed in 4% formaline (pH 7.4), embedded in paraffin followed by cutting with a microtome (3 µm thickness). The slides were deparaffinized in xylol for 20 minutes and then rehydrated in descending series of ethanol (100%, 100%, 96%, 96%, 70%, and 70%). For antigen retrieval the slides were boiled in citrate buffer (pH 6.0) for 40 min, and then allowed to cool down for 15 min. After washing with PBS buffer the endogenous peroxidase was blocked with H2O2 for 15 min at room temperature. After washing in PBS the slides were incubated with the antibody against PAX2 (dilution 1∶100) for 60 min at room temperature and washed in PBS again.

    The secondary antibody was incubated for 20 min at room temperature and after washing the slides in PBS the biotin streptavidine label was incubated for 20 min at room temperature. A detection kit including horseradish peroxidase and diaminobenzidine as chromogene was applied for 5 min. Counterstaining was performed with hematoxilin for 6 min.

    (A) In normal sweat glands, Pax2 is expressed in gland epithelial cells (black arrows) while intermingled stromal cells only show very weak or absent nuclear Pax2 expression (green arrow) Bar represents 100 µm.

    (B, C) Normal appearing epidermal cell layers adjacent to (B, bar represent 100 µm) nevi or (C, bar represent 200 µm) malignant melanoma show a differentially Pax2 expression with strongest Pax2 levels in germinal basal cell layers (black arrows) decreasing in higher differentiated keratinocytes and finally being absent in corneocytes (green arrows).

    (D) Malignant melanoma cells - heterogeneous nuclear Pax2 expression. Strongest expression in large atypical nuclei with prominent nucleoli (black arrows). 

    (E, F) Pax2 expression in intradermal nevi was heterogeneous and did not correlate with histological features.

  • Immunohistochemical analysis of Human malignant melanoma tissue, staining Pax2 with unpurified ab79389.
    Antigen retrieval was performed via heat mediation in a citrate buffer (pH 6). Sections were blocked with 0.1% Triton X-100/PBS containing 1% BSA and 10% horse serum for 1 hour. Samples were incubated with primary antibody overnight at 4°C. A Cy3®-conjugated goat anti-rabbit IgG was used as the secondary antibody.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of (A) human fetal kidney, (B) human normal kidney and (C) human renal cell carcinoma tissues labelled Pax2 with unpurified ab79389 at a dilution of 1/1000.

  • Immunocytochemistry/Immunofluorescence analysis of HEK293 cells labelling Pax2 with purified ab79389 at a dilution of 1/50. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/50) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

  • Anti-Pax2 antibody [EP3251] (ab79389) at 1/1000 dilution (purified) + Human fetal kidney tissue lysate at 20 µg

    Secondary
    HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 45 kDa
    Observed band size: 45 kDa



    Blocking and dilution buffer: 5% NFDM/TBST
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling Pax2 with purified ab79389 at a dilution of 1/500. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue labelling Pax2 with purified ab79389 at a dilution of 1/500. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cerebral cortex tissue labelling Pax2 with purified ab79389 at a dilution of 1/500. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Flow Cytometry analysis of K562 cells labelling Pax2 with purified ab79389 at a dilution of 1/50 (red). Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Anti-Pax2 antibody [EP3251] (ab79389) at 1/10000 dilution (unpurified) + Human fetal kidney tissue lysate at 10 µg

    Secondary
    HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

    Predicted band size: 45 kDa
    Observed band size: 45 kDa

  • Overlay histogram showing HEK293 cells stained with unpurified ab79389 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (unpurified ab79389, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a significantly decreased signal in HEK293 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

参考文献

This product has been referenced in:
  • Jahangiri R  et al. PAX2 expression is correlated with better survival in tamoxifen-treated breast carcinoma patients. Tissue Cell 52:135-142 (2018). IHC-P ; Human . Read more (PubMed: 29857823) »
  • Xu Q  et al. Renal carcinoma/kidney progenitor cell chimera organoid as a novel tumorigenesis gene discovery model. Dis Model Mech 10:1503-1515 (2017). Read more (PubMed: 29084770) »

See all 18 Publications for this product

レビューと Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Cat Tissue sections (Brain)
Antigen retrieval step
None
Permeabilization
Yes - Tween20
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

投稿 Aug 06 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer (PH6)
Permeabilization
No
Specification
kidney
Blocking step
x0909 (DAKO blocking product) as blocking agent for 15 minute(s) · Concentration: 100% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 May 29 2017

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 35 minute(s) · Concentration: 5% · Temperature: 20°C
Sample
Human Cell (differentiated iPSC)
Specification
differentiated iPSC
Permeabilization
Yes - Triton X
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Feb 26 2015

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (4-12% acrylamide)
Sample
Human Cell lysate - whole cell (Renal cancer cell line)
Specification
Renal cancer cell line
Blocking step
Li-cor Blocking Buffer PBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Jul 31 2014



According to our records we have not received a customer report with similar results.
You have tried quite a few optimizations, but the result is still not satisfying.

Could you please let me know the species of your samples?
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Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

The details you have kindly provided will enable us to investigate this...

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Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

I a...

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Thank you again for your email.
The concentration of your lot is 0.2290 mg/ml.

Please let me know if you have anymore questions.

In order to help you, I would need to know which lot./ batch number you received from us?

I am looking forward to your reply.

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Breast)
Specification
Breast
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TRIS-ETDA-Buffer pH 9,0
Permeabilization
Yes - Wash Buffer with Tween from Dako
Username

Mr. Rudolf Jung

Verified customer

投稿 Sep 28 2012

1-10 of 13 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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