Recombinant
RabMAb

Recombinant Anti-WDR77 antibody [EPR10708(B)] (ab154190)

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Overview

  • Product name

    Anti-WDR77 antibody [EPR10708(B)]
    See all WDR77 primary antibodies

  • Description

    Rabbit monoclonal [EPR10708(B)] to WDR77

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cytmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide corresponding to residues in Human WDR77 (UniProt ID: Q9BQA1).

  • Positive control

    • HeLa, LnCAP, 293 and fetal heart lysates; Human colon and prostate tissues.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Applications

Our Abpromise guarantee covers the use of ab154190 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 1/1000 - 1/10000. Predicted molecular weight: 37 kDa.
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF 1/100 - 1/250.
IP 1/10 - 1/100.
Flow Cyt 1/100 - 1/10000.

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

 

Target

  • Function

    Non-catalytic component of the 20S PRMT5-containing methyltransferase complex, which modifies specific arginines to dimethylarginines in several spliceosomal Sm proteins. This modification targets Sm proteins to the survival of motor neurons (SMN) complex for assembly into small nuclear ribonucleoprotein core particles. Might play a role in transcription regulation. The 20S PRMT5-containing methyltransferase complex also methylates the Piwi proteins (PIWIL1, PIWIL2 and PIWIL4), methylation of Piwi proteins being required for the interaction with Tudor domain-containing proteins and subsequent localization to the meiotic nuage.

  • Tissue specificity

    Highly expressed in heart, skeletal muscle, spleen, testis, uterus, prostate and thymus. In testis, expressed in germ cells and Leydig cells, but not in peritubular myocytes, nor in Sertoli cells. Expressed in prostate cancers, in seminomas and in Leydig cell tumors.

  • Sequence similarities

    Contains 5 WD repeats.

  • Developmental stage

    Expressed in Leydig cells during fetal testicular development, especially during the second semester. Germ cells expression is detected as early as 10 weeks of gestation.

  • Cellular localization

    Nucleus. Cytoplasm. Nuclear in Leydig cells and cytoplasmic in germ cells during fetal testicular development. In adult testis, predominantly nuclear. Subcellular location varies from nuclear to cytoplasmic in various tumors.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • 2610312E17Rik antibody
    • Androgen receptor cofactor p44 antibody
    • C79984 antibody
    • HKMT1069 antibody
    • MEP 50 antibody
    • MEP-50 antibody
    • MEP50 antibody
    • MEP50_HUMAN antibody
    • Methylosome protein 50 antibody
    • MGC2722 antibody
    • Nbla10071 antibody
    • p44 antibody
    • p44/Mep50 antibody
    • RGD1310479 antibody
    • RP11 552M11.3 antibody
    • WD repeat containing protein 77 antibody
    • WD repeat domain 77 antibody
    • WD repeat-containing protein 77 antibody
    • WDR77 antibody
    see all

Images

  • Western blot - Anti-WDR77 antibody [EPR10708(B)] (ab154190)
    Western blot - Anti-WDR77 antibody [EPR10708(B)] (ab154190)
    All lanes : Anti-WDR77 antibody [EPR10708(B)] (ab154190) at 1/1000 dilution

    Lane 1 : HeLa cell lysate
    Lane 2 : LnCAP cell lysate
    Lane 3 : 293T cell lysate
    Lane 4 : fetal heart lysate

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 37 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-WDR77 antibody [EPR10708(B)] (ab154190)
    Immunocytochemistry/ Immunofluorescence - Anti-WDR77 antibody [EPR10708(B)] (ab154190)

    Immunocytochemistry/Immunofluorescence analysis of HT-29 (Human colorectal adenocarcinoma cell line) labeling WDR77 with Purified ab154190 at 1/500 dilution (5 µg/ml). Cells were fixed with 4% PFA and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) at 1/1000 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

  • Flow Cytometry - Anti-WDR77 antibody [EPR10708(B)] (ab154190)
    Flow Cytometry - Anti-WDR77 antibody [EPR10708(B)] (ab154190)
    Overlay histogram showing HeLa cells stained with ab154190 (red line). The cells were fixed with 80% 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 (ab154190, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
  • Immunocytochemistry/ Immunofluorescence - Anti-WDR77 antibody [EPR10708(B)] (ab154190)
    Immunocytochemistry/ Immunofluorescence - Anti-WDR77 antibody [EPR10708(B)] (ab154190)This image is courtesy of an Abreview submitted by Melanie Blasius

    ab154190 staining WRD77 (green) in U2OS cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.2% Triton X-100 in PBS. Samples were incubated with primary antibody (1/150 in PBS + 5% BSA) for 1 hour at 20°C. An Alexa Fluor®488-conjugated Goat anti-rabbit IgG polyclonal (1/800) was used as the secondary antibody. Red - DAPI.

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WDR77 antibody [EPR10708(B)] (ab154190)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WDR77 antibody [EPR10708(B)] (ab154190)

    Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling WDR77 with ab154190 at 1/50 dilution

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WDR77 antibody [EPR10708(B)] (ab154190)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WDR77 antibody [EPR10708(B)] (ab154190)

    Immunohistochemical analysis of paraffin-embedded Human prostate tissue labeling WDR77 with ab154190 at 1/50 dilution.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WDR77 antibody [EPR10708(B)] (ab154190)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WDR77 antibody [EPR10708(B)] (ab154190)

    Immunohistochemical analysis of paraffin embedded Human thyroid gland tissue using ab154190 showing +ve staining.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WDR77 antibody [EPR10708(B)] (ab154190)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WDR77 antibody [EPR10708(B)] (ab154190)

    Immunohistochemical analysis of paraffin embedded Human normal testis tissue using ab154190 showing +ve staining.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WDR77 antibody [EPR10708(B)] (ab154190)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WDR77 antibody [EPR10708(B)] (ab154190)

    Immunohistochemical analysis of paraffin embedded Human breast carcinoma tissue using ab154190 showing +ve staining.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WDR77 antibody [EPR10708(B)] (ab154190)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WDR77 antibody [EPR10708(B)] (ab154190)

    Immunohistochemical analysis of paraffin embedded Human thyroid gland carcinoma tissue using ab154190 showing +ve staining.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

References

This product has been referenced in:

  • Qi H  et al. Sirtuin 7-mediated deacetylation of WD repeat domain 77 (WDR77) suppresses cancer cell growth by reducing WDR77/PRMT5 transmethylase complex activity. J Biol Chem 293:17769-17779 (2018). Read more (PubMed: 30282801) »
  • Lubyova B  et al. PRMT5: A novel regulator of Hepatitis B virus replication and an arginine methylase of HBV core. PLoS One 12:e0186982 (2017). WB ; Human . Read more (PubMed: 29065155) »
See all 2 Publications for this product

Publishing research using ab154190? Please let us know so that we can cite the reference in this datasheet.

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Immunocytochemistry/ Immunofluorescence abreview for Anti-WDR77 antibody [EPR10708(B)]

 Good
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (U2OS cells)
Specification
U2OS cells
Permeabilization
Yes - PBS+ 0.2% Triton-X 100, 10 minutes at RT
Fixative
Paraformaldehyde
Read More

Dr. Melanie Blasius

Verified customer

Submitted Jun 28 2013

Western blot

 Excellent
Abreviews
abreview image
Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing (NuPAGE® Novex® 3-8% Tris-Acetate Gel 1.0 mm)
Sample
Human Cell lysate - whole cell (U2OS cells)
Specification
U2OS cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Read More

Dr. Melanie Blasius

Verified customer

Submitted May 20 2013

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