Product Info Summary
SKU: | A31776 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Rabbit |
Application: | ELISA, Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-VPRBP/DCAF1 Antibody Picoband®
SKU/Catalog Number
A31776
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-VPRBP/DCAF1 Antibody Picoband® catalog # A31776. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-VPRBP/DCAF1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A31776)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human VPRBP/DCAF1 recombinant protein (Position: M1-R195).
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A31776 is reactive to DCAF1 in Human, Mouse, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
180 kDa
Calculated molecular weight
14377 MW
Background of DCAF1
VprBP was first identified as a protein that can interact with HIV-1 viral protein R. It is a component of the CUL4A-RBX1-DDB1-VprBP/DCAF1 E3 ubiquitin-protein ligase complex that could interact with HIV-1 virus Vpr protein and HIV-2 virus Vpx protein. VprBP is a 1,507-amino acid protein that contains conserved domains, including YXXY repeats, the Lis homology motif, and WD40 repeats. Through binding to Vpr, VprBP allows Vpr to modulate the catalytic activity of the CUL4-DDB1 complex, which in turn leads to the induction of G2 phase arrest in the virus-infected cells. Recently it has been reported that VprBP is able to regulate the p53-induced transcription and apoptotic pathway.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A31776 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.25-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, Human
Positive Control
WB: human HEK293 whole cell, human HepG2 whole cell, human K562 whole cell, rat testis tissue, mouse testis tissue
IHC: human placenta tissue, human breast cancer tissue, human ovarian adenocarcinoma tissue, human pancreatic cancer tissue, human rectal cancer tissue, human gastric cancer tissue
ICC/IF: PC-3 cell
FCM: U251 cell
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of VPRBP/DCAF1 using anti-VPRBP/DCAF1 antibody (A31776).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30ug of sample under reducing conditions.
Lane 1: human HEK293 whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: rat testis tissue lysates,
Lane 5: mouse testis tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-VPRBP/DCAF1 antigen affinity purified polyclonal antibody (Catalog # A31776) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for VPRBP/DCAF1 at approximately 180KD. The expected band size for VPRBP/DCAF1 is at 180KD.
Click image to see more details
Figure 2. IHC analysis of VPRBP/DCAF1 using anti-VPRBP/DCAF1 antibody (A31776).
VPRBP/DCAF1 was detected in paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-VPRBP/DCAF1 Antibody (A31776) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 3. IHC analysis of VPRBP/DCAF1 using anti-VPRBP/DCAF1 antibody (A31776).
VPRBP/DCAF1 was detected in paraffin-embedded section of human breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-VPRBP/DCAF1 Antibody (A31776) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 4. IHC analysis of VPRBP/DCAF1 using anti-VPRBP/DCAF1 antibody (A31776).
VPRBP/DCAF1 was detected in paraffin-embedded section of human ovarian adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-VPRBP/DCAF1 Antibody (A31776) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 5. IHC analysis of VPRBP/DCAF1 using anti-VPRBP/DCAF1 antibody (A31776).
VPRBP/DCAF1 was detected in paraffin-embedded section of human pancreatic cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-VPRBP/DCAF1 Antibody (A31776) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 6. IHC analysis of VPRBP/DCAF1 using anti-VPRBP/DCAF1 antibody (A31776).
VPRBP/DCAF1 was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-VPRBP/DCAF1 Antibody (A31776) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 7. IHC analysis of VPRBP/DCAF1 using anti-VPRBP/DCAF1 antibody (A31776).
VPRBP/DCAF1 was detected in paraffin-embedded section of human gastric cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-VPRBP/DCAF1 Antibody (A31776) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 8. IF analysis of VPRBP/DCAF1 using anti-VPRBP/DCAF1 antibody (A31776).
VPRBP/DCAF1 was detected in immunocytochemical section of PC-3 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-VPRBP/DCAF1 Antibody (A31776) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Click image to see more details
Figure 9. Flow Cytometry analysis of U251 cells using anti-VPRBP/DCAF1 antibody (A31776).
Overlay histogram showing U251 cells stained with A31776 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-VPRBP/DCAF1 Antibody (A31776, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Protein Target Info & Infographic
Gene/Protein Information For DCAF1 (Source: Uniprot.org, NCBI)
Gene Name
DCAF1
Full Name
DDB1- and CUL4-associated factor 1
Weight
14377 MW
Superfamily
VPRBP/DCAF1 family
Alternative Names
Protein Bop; BH3-only protein; Retrotransposon Gag-like protein 10; RTL10; BOP, C22orf29 DCAF1 RIP, VPRBP DDB1 and CUL4 associated factor 1 DDB1- and CUL4-associated factor 1|HIV-1 Vpr-binding protein|Vpr (HIV-1) binding protein|Vpr-binding protein|protein VPRBP|serine/threonine-protein kinase VPRBP|vpr-interacting protein
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on DCAF1, check out the DCAF1 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for DCAF1: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-VPRBP/DCAF1 Antibody Picoband® (A31776)
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