UAF1/WDR48 Antibody - BSA Free
Novus Biologicals | Catalog # NBP1-81404
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Key Product Details
Validated by
Independent Antibodies
Species Reactivity
Validated:
Human
Predicted:
Mouse (100%), Rat (100%). Backed by our 100% Guarantee.
Applications
Validated:
Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence
Cited:
Western Blot, Immunoprecipitation, Chemotaxis
Label
Unconjugated
Antibody Source
Polyclonal Rabbit IgG
Format
BSA Free
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Product Specifications
Immunogen
This antibody was developed against Recombinant Protein corresponding to amino acids: SIIQCHILNDKRHILTKDTNNNVAYWDVLKACKVEDLGKVDFEDEIKKRFKMVYVPNWFSVDLKTGMLTITLDESDCFAAWVSAKDAGF
Clonality
Polyclonal
Host
Rabbit
Isotype
IgG
Theoretical MW
76 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Scientific Data Images for UAF1/WDR48 Antibody - BSA Free
Immunocytochemistry/ Immunofluorescence: UAF1/WDR48 Antibody [NBP1-81404]
Immunocytochemistry/Immunofluorescence: UAF1/WDR48 Antibody [NBP1-81404] - Staining of human cell line A-431 shows localization to vesicles. Antibody staining is shown in green.Immunohistochemistry-Paraffin: UAF1/WDR48 Antibody [NBP1-81404]
Immunohistochemistry-Paraffin: UAF1/WDR48 Antibody [NBP1-81404] - Staining of human cervix, uterine shows moderate cytoplasmic positivity in squamous epithelial cells.Immunohistochemistry-Paraffin: UAF1/WDR48 Antibody [NBP1-81404]
Immunohistochemistry-Paraffin: UAF1/WDR48 Antibody [NBP1-81404] - Staining of human testis shows moderate to strong positivity.Immunohistochemistry-Paraffin: UAF1/WDR48 Antibody [NBP1-81404]
Immunohistochemistry-Paraffin: UAF1/WDR48 Antibody [NBP1-81404] - Staining of human endometrium shows moderate cytoplasmic positivity in glandular cells.Immunohistochemistry-Paraffin: UAF1/WDR48 Antibody [NBP1-81404]
Immunohistochemistry-Paraffin: UAF1/WDR48 Antibody [NBP1-81404] - Staining of human cerebral cortex shows cytoplasmic positivity in neuronal cells.Simple Western: UAF1/WDR48 Antibody [NBP1-81404]
Simple Western: UAF1/WDR48 Antibody [NBP1-81404] - Simple Western lane view shows a specific band for UAF1/WDR48 in 0.2 mg/ml of RT-4 (Left) and U-251MG (Right) lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.Simple Western: UAF1/WDR48 Antibody [NBP1-81404]
Simple Western: UAF1/WDR48 Antibody [NBP1-81404] - Electropherogram image(s) of corresponding Simple Western lane view. UAF1/WDR48 antibody was used at 1:20 dilution on RT-4 and U-251MG lysate(s).Western Blot: UAF1/WDR48 Antibody [NBP1-81404] -
Western Blot: UAF1/WDR48 Antibody [NBP1-81404] - EBNA3C associates with the WDR48/USP46 complex in EBNA3C-F-HA LCLs.Immunoprecipitation assay using Flag agarose to retrieve protein complexes from EBNA3C-F-HA LCLs (E3C-F-HA) is compared to flag immunoprecipiates from untagged wildtype (WT) LCLs. One percent of total cell lysate (Input) or immunoprepicitated specimens using Flag agarose (Flag IP) were separated by SDS PAGE & probed using antibodies to EBNA3C, RBPJ, CtBP1, WDR48, WDR20, USP46, or NF-kB p65. Image collected & cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.ppat.1004822), licensed under a CC-BY license. Not internally tested by Novus Biologicals.Western Blot: UAF1/WDR48 Antibody [NBP1-81404] -
Western Blot: UAF1/WDR48 Antibody [NBP1-81404] - Deletion of EBNA3A residues 920–944 disrupts WDR48 binding without affecting CtBP1 association.Co-immunoprecipitation assay to assess binding of EBNA3A mutants to WDR48 (A) & CtBP1 (B). For these assays, flag tagged full length EBNA3A (1–944), an EBNA3A CtBP1 binding mutant (mCtBP), an EBNA3A mutant lacking the C-terminal 25 residues (1–919), or vector control (pSG5) was co-transfected with Xpress-WDR48 or HA-CtBP1. Lysates were immunoprecipitated with Flag agarose (A) or HA agarose (B), separated by SDS PAGE, & probed with WDR48, RBPJ, flag, EBNA3A & HA antibodies. Image collected & cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.ppat.1004822), licensed under a CC-BY license. Not internally tested by Novus Biologicals.Western Blot: UAF1/WDR48 Antibody [NBP1-81404] -
Western Blot: UAF1/WDR48 Antibody [NBP1-81404] - Identification of EBNA3A & EBNA3C domains that mediate WDR48 association.Immunoprecipitation assays to map WDR48 binding regions within EBNA3A (A) & EBNA3C (B). 293T cells were co-transfected with Xpress tagged WDR48 & flag tagged full length EBNA3A, EBNA3C, or the indicated EBNA3A or EBNA3C deletion mutants. Cell lysates were immunoprecipitated with Flag agarose, separated by SDS PAGE, & probed for WDR48 (anti-Xpress) & EBNA3 proteins (anti-Flag). (C) Comparison of WDR48 binding results (from B) with previously published RBPJ binding results & LCL growth phenotype for each EBNA3C mutant [21]. Image collected & cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.ppat.1004822), licensed under a CC-BY license. Not internally tested by Novus Biologicals.Western Blot: UAF1/WDR48 Antibody [NBP1-81404] -
Western Blot: UAF1/WDR48 Antibody [NBP1-81404] - EBNA3 proteins preferentially bind the WDR48 subunit of the USP46 DUB complex.(A) Immunoprecipitation assay in 293T cells demonstrating association of flag tagged EBNA3 proteins (F-E3A, F-E3B, & F-E3C) with WDR48 (left) & WDR20 (right). (B) Immunoprecipitation assay demonstrating WDR48 cotransfection enhanced USP46 association with EBNA3s (right panel, compare lanes 3–5 with 8–10). Epitope tagged BNRF1 (F-HA-BNRF1), an EBV protein of approximately the same size as the EBNA3 is included as an additional negative control. One percent (panel A) or two percent (panel B) of the input are shown for comparison. Image collected & cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.ppat.1004822), licensed under a CC-BY license. Not internally tested by Novus Biologicals.Western Blot: UAF1/WDR48 Antibody [NBP1-81404] -
Western Blot: UAF1/WDR48 Antibody [NBP1-81404] - WDR48 coimmunoprecipitates with RBPJ in EBV infected cells.Co-immunoprecipitation assays comparing the association of RBPJ with WDR48 in LCLs with that observed in EBV negative BL41 cells. Cell lysates were immunoprecipitated with polyclonal RBPJ sera, separated by SDS PAGE, & probed for EBNA3A, EBNA3C, WDR48, & RBPJ (as indicated). Image collected & cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.ppat.1004822), licensed under a CC-BY license. Not internally tested by Novus Biologicals.Western Blot: UAF1/WDR48 Antibody [NBP1-81404] -
Western Blot: UAF1/WDR48 Antibody [NBP1-81404] - ChIP assay for WDR48 at the p14ARF promoter.Chromatin immunoprecipitation (ChIP) assays were performed using antibodies for WDR48 (A) from EBNA3C-HT LCLs that were grown in the presence of 4HT (dark gray) or after 14 days of growth in the absence of 4HT (light gray). Amount of genomic DNA was measured by real time PCR using primers specific to the EBNA3C binding site in the p14ARF promoters or sites near the EIF2AK3 & PPIA genes which bind cell transcription factors but not EBNA3C. The bar graph represents the amount of DNA precipitated relative to the amount of DNA in the corresponding input sample. The experiment shown is representative of four independent experiments & error bars indicating standard error of the mean within this experiment. Asterisk denotes that the difference in ChIP signal seen at the p14ARF promoter is statistically significant (p = 0.01). (B) Western blot for USP46, WDR48, & tubulin levels in whole cell lysates from EBNA3CHT LCLs grown in the presence of 4HT or after 14 days of growth in the absence of 4HT. Image collected & cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.ppat.1004822), licensed under a CC-BY license. Not internally tested by Novus Biologicals.Western Blot: UAF1/WDR48 Antibody [NBP1-81404] -
Western Blot: UAF1/WDR48 Antibody [NBP1-81404] - WDR48 SLD2 mediates binding to EBNA3B & EBNA3C, but is not required for EBNA3A binding.Immunoprecipitation assays were performed to assess effect of deleting the WDR48 SUMO-like domains (SLDs) on EBNA3 binding. (A) 293T cells were co-transfected with vector control, Xpress tagged full length WDR48 (FL) or a deletion mutant lacking the SLD2 domain (WDR48 1–634) & flag tagged EBNA3A 1–944 (F-E3A), EBNA3B 394–938 (F-E3B) or EBNA3C 365–545 (F-E3C). Cell lysates were immunoprecipitated with Flag agarose, separated by SDS PAGE, & probed with WDR48 or Flag antibody. (B) Immunoprecipitation assay to determine effect of deleting SLD1/2 on WDR48 binding to EBNA3A. Assays were performed as describe above with co-transfection of EBNA3A WT & either full length WDR48 (FL), WDR48 1–535, or WDR48 1–430 (which lacks both SLD1 & SLD2). Image collected & cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.ppat.1004822), licensed under a CC-BY license. Not internally tested by Novus Biologicals.Applications for UAF1/WDR48 Antibody - BSA Free
Application
Recommended Usage
Immunocytochemistry/ Immunofluorescence
0.25-2 ug/ml
Immunohistochemistry
1:50 - 1:200
Immunohistochemistry-Paraffin
1:50 - 1:200
Western Blot
0.04-0.4 ug/ml
Application Notes
For IHC-Paraffin, HIER pH 6 retrieval is recommended. ICC/IF Fixation Permeabilization: Use PFA/Triton X-100.
In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in RT-4 and U-251MG, separated by Size, antibody dilution of 1:20, apparent MW was 87 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.
In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in RT-4 and U-251MG, separated by Size, antibody dilution of 1:20, apparent MW was 87 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.
Formulation, Preparation, and Storage
Purification
Affinity purified
Formulation
PBS (pH 7.2) and 40% Glycerol
Format
BSA Free
Preservative
0.02% Sodium Azide
Concentration
Concentrations vary lot to lot. See vial label for concentration. If unlisted please contact technical services.
Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Background: UAF1
Long Name
USP1-associated Factor 1
Alternate Names
P80, WDR48
Gene Symbol
WDR48
UniProt
Additional UAF1 Products
Product Documents for UAF1/WDR48 Antibody - BSA Free
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Product Specific Notices for UAF1/WDR48 Antibody - BSA Free
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
Related Research Areas
Citations for UAF1/WDR48 Antibody - BSA Free
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Antigen Retrieval Protocol (PIER)
- Antigen Retrieval for Frozen Sections Protocol
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- Chromogenic IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Chromogenic Immunohistochemistry Staining of Frozen Tissue
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- Fluorescent IHC Staining of Frozen Tissue Protocol
- Graphic Protocol for Heat-induced Epitope Retrieval
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Graphic Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- IHC Sample Preparation (Frozen sections vs Paraffin)
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunofluorescent IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Immunohistochemistry Frozen Troubleshooting
- Immunohistochemistry Paraffin Troubleshooting
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for Heat-Induced Epitope Retrieval (HIER)
- Protocol for Making a 4% Formaldehyde Solution in PBS
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation & Fixation of Cells on Coverslips
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections - Graphic
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Immunohistochemistry
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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