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Product Info Summary
| SKU: | A08280-2 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-Apc5/ANAPC5 Antibody Picoband®
SKU/Catalog Number
A08280-2
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Apc5/ANAPC5 Antibody Picoband® catalog # A08280-2. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human. 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-Apc5/ANAPC5 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A08280-2)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.01mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human Apc5/ANAPC5 recombinant protein (Position: M1-D266).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A08280-2 is reactive to ANAPC5 in Human
Observed Molecular Weight
85 kDa
Calculated molecular weight
85.1 kDa
Background of ANAPC5
Anaphase-promoting complex subunit 5 is an enzyme that in humans is encoded by the ANAPC5 gene. This gene encodes a tetratricopeptide repeat-containing component of the anaphase promoting complex/cyclosome (APC/C), a large E3 ubiquitin ligase that controls cell cycle progression by targeting a number of cell cycle regulators such as B-type cyclins for 26S proteasome-mediated degradation through ubiquitination. The encoded protein is required for the proper ubiquitination function of APC/C and for the interaction of APC/C with transcription coactivators. It also interacts with polyA binding protein and represses internal ribosome entry site-mediated translation. Multiple transcript variants encoding different isoforms have been found for this gene. These differences cause translation initiation at a downstream AUG and result in a shorter protein (isoform b), compared to isoform a.
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
A08280-2 is guaranteed for ELISA, Flow Cytometry, IF, 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
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: human HL-60 whole cell, human HEK293 whole cell, human K562 whole cell, human Jurkat whole cell, human Raji whole cell
ICC/IF: A431 cell
FCM: 293T cell
Validation Images & Assay Conditions
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Western blot analysis of Apc5/ANAPC5 using anti-Apc5/ANAPC5 antibody (A08280-2).
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 50ug of sample under reducing conditions.
Lane 1: human HL-60 whole cell lysates,
Lane 2: human HEK293 whole cell lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: human Jurkat whole cell lysates,
Lane 5: human Raji whole cell 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-Apc5/ANAPC5 antigen affinity purified polyclonal antibody (Catalog # A08280-2) 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 Apc5/ANAPC5 at approximately 85KD. The expected band size for Apc5/ANAPC5 is at 85KD.
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IF analysis of Apc5/ANAPC5 using anti-Apc5/ANAPC5 antibody (A08280-2).
Apc5/ANAPC5 was detected in immunocytochemical section of A431 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-Apc5/ANAPC5 Antibody (A08280-2) 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.
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Flow Cytometry analysis of 293T cells using anti-Apc5/ANAPC5 antibody (A08280-2).
Overlay histogram showing 293T cells stained with A08280-2 (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-Apc5/ANAPC5 Antibody (A08280-2, 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.
Specific Publications For Anti-Apc5/ANAPC5 Antibody Picoband® (A08280-2)
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