Anti-Apc5/ANAPC5 Antibody Picoband™

Product Name

Anti-Apc5/ANAPC5 Antibody Picoband™

View all Apc5 Antibodies

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.

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).

*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

A08280-2 is reactive to ANAPC5 in Human

Applications

A08280-2 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

85 kDa

Calculated molecular weight

85.077kDa

Background of Apc5

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.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

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, 1-3μg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5μg/ml, Human

Validation Images & Assay Conditions

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Figure 1. 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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Figure 2. 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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Figure 3. Flow Cytometry analysis of 293T cells using anti-Apc5/ANAPC5 antibody (A08280-2).
Overlay histogram showing 293T cells stained with A08280-2 (Blue line).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 (Red line) was also used as a control.

Gene/Protein Information For ANAPC5 (Source: Uniprot.org, NCBI)

Alternative Names

anaphase promoting complex subunit 5; anaphase-promoting complex subunit 5; APC5Cyclosome subunit 5 ANAPC5 APC5 anaphase promoting complex subunit 5 anaphase-promoting complex subunit 5|cyclosome subunit 5

For more info on ANAPC5, check out the ANAPC5 Infographic

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In this infographic, you will see the following information for ANAPC5: 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].

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