Anti-CDCA8 Antibody Picoband™

Product Name

Anti-CDCA8 Antibody Picoband™

View all Borealin Antibodies

SKU/Catalog Number

A06612-2

Size

100 µg/vial

Form

Lyophilized

Description

Boster Bio Anti-CDCA8 Antibody Picoband™ catalog # A04479-2. Tested in WB, ICC/IF, FCM, ELISA applications. This antibody reacts with Human.

Storage & Handling

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 freezing and thawing.

Cite This Product

Anti-CDCA8 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A06612-2)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.

Clonality

Polyclonal

Isotype

IgG

Immunogen

E.coli-derived human CDCA8 recombinant protein (Position: D72-K280). Human CDCA8 shares 76% and 77.8% amino acid (aa) sequence identity with mouse and rat CDCA8, respectively.

*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

A06612-2 is reactive to Cdca8 in Human

Applications

A06612-2 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

35 kDa

Calculated molecular weight

52588 MW

Background of Borealin

Borealin is a protein that in humans is encoded by the CDCA8 gene. This gene encodes a component of the chromosomal passenger complex. This complex is an essential regulator of mitosis and cell division. This protein is cell-cycle regulated and is required for chromatin-induced microtubule stabilization and spindle formation. Alternate splicing results in multiple transcript variants. Pseudgenes of this gene are found on chromosomes 7, 8 and 16.

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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Reconstitution

Adding 0.2 ml of distilled water will yield a concentration of 500 µg/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, 0.25-0.5 µg/ml, Human
Immunocytochemistry/Immunofluorescence, 5 µg/ml, Human
Flow Cytometry (Fixed), 1-3 µg/1x10⁶ cells, Human
ELISA, 0.1-0.5 µg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of CDCA8 using anti-CDCA8 antibody (A06612-2).
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 30 ug of sample under reducing conditions.
Lane 1: human K562 whole cell lysates,
Lane 2: human HEL whole cell lysates,
Lane 3: human A431 whole cell lysates,
Lane 4: human A549 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-CDCA8 antigen affinity purified polyclonal antibody (Catalog # A06612-2) 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 CDCA8 at approximately 35 kDa. The expected band size for CDCA8 is at 31 kDa.

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Figure 2. IF analysis of CDCA8 using anti-CDCA8 antibody (A06612-2) and anti-Beta Tubulin antibody (M01857-3).
CDCA8 was detected in immunocytochemical section of U2OS cell. 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 5 μg/mL rabbit anti-CDCA8 Antibody (A06612-2) and mouse anti-Beta Tubulin antibody (M01857-3) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) and DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Figure 3. Flow Cytometry analysis of K562 cells using anti-CDCA8 antibody (A06612-2).
Overlay histogram showing K562 cells stained with A06612-2 (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-CDCA8 Antibody (A06612-2, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For Cdca8 (Source: Uniprot.org, NCBI)

Alternative Names

BOR; BOREALIN; cell division cycle associated 8; Cell division cycle-associated protein 8; Dasra B; DasraB; dasra-B; FLJ10468; FLJ12042; hDasra-B; MESRGP; PESCRG3; Pluripotent embryonic stem cell-related gene 3 protein Cdca8|4831429J16Rik, AU044747, BOR, Bo, Borealin, D4Ertd421, D4Ertd421e, Da, DasraB, MESRGP|cell division cycle associated 8|borealin|MESrg|cell division cycle-associated protein 8|embryonic stem cell-related

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In this infographic, you will see the following information for Cdca8: 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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