Anti-SIAH Interacting Protein/CACYBP Antibody

Product Name

Anti-SIAH Interacting Protein/CACYBP Antibody

View all Cacybp Antibodies

SKU/Catalog Number

PA1759

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-SIAH Interacting Protein/CACYBP Antibody catalog # PA1759. Tested in IP, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.

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-SIAH Interacting Protein/CACYBP Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1759)

Host

Rabbit

Contents

Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.01mg Thimerosal, 0.01mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

A synthetic peptide corresponding to a sequence in the middle region of human CACYBP, identical to the related rat and mouse sequences.

*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

PA1759 is reactive to Cacybp in Human, Mouse, Rat

Reconstitution

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

Observed Molecular Weight

27 kDa

Calculated molecular weight

26210 MW

Background of Cacybp

CACYBP (Calcyclin-binding protein), also called SIP, is a protein that in humans is encoded by the CACYBP gene. The full-length SIP cDNA encodes a predicted 228-amino acid protein. Sequence analysis of the shortest cDNA derived by 2-hybrid screening revealed an 8-amino acid difference in the deduced open reading frame followed by a stop codon, resulting in a predicted 80-amino acid protein, SIP-short (SIPS). The CACYBP gene is mapped on 1q25.1. It may be involved in calcium-dependent ubiquitination and subsequent proteosomal degradation of target proteins. It probably serves as a molecular bridge in ubiquitin E3 complexes and participates in the ubiquitin-mediated degradation of beta-catenin. Two alternatively spliced transcript variants encoding different isoforms have been found for this gene. The C-terminal region of SIP that is homologous to SGT1 was able to complement defects in yeast strains containing SGT1 mutant alleles, demonstrating conservation of SGT1 and SIP protein function.

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.

View more details

Applications

PA1759 is guaranteed for Flow Cytometry, IP, 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, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry(Fixed), 1-3 μg/1x10⁶ cells, Human
Immunoprecipitation, 0.5-2 μg/ml, Human

Positive Control

Validation Images & Assay Conditions

Click image to see more details

Figure 1. Western blot analysis of CACYBP using anti-CACYBP antibody (PA1759).
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 MCF-7 whole cell lysates,
Lane 2: human RT4 whole cell lysates,
Lane 3: human SW620 whole cell lysates,
Lane 4: human U251 whole cell lysates,
Lane 5: rat liver tissue lysates,
Lane 6: rat brain tissue lysates,
Lane 7: mouse liver tissue lysates,
Lane 8: mouse brain tissue 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-CACYBP antigen affinity purified polyclonal antibody (Catalog # PA1759) 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 CACYBP at approximately 27 kDa. The expected band size for CACYBP is at 27 kDa.

Click image to see more details

Figure 4. Immunoprecipitating CACYBP in U251 whole cell lysate.
Western blot analysis of CACYBP using anti-CACYBP antibody (PA1759).
Lane 1: U251 whole cell lysates (30ug),
Lane 2: Rabbit control IgG instead of anti-CACYBP antibody in U251 whole cell lysate,
Lane 3: anti-CACYBP antibody (2μg) + U251 whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-CACYBP antigen affinity purified polyclonal antibody (PA1759) at a dilution of 0.5 μg/mL and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1197). A specific band was detected for CACYBP at approximately 27 kDa. The expected band size for CACYBP is at 27 kDa.

Click image to see more details

Figure 2. IF analysis of CACYBP using anti-CACYBP antibody (PA1759).
CACYBP was detected in an immunocytochemical section of Hela 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 5 μg/mL rabbit anti-CACYBP Antibody (PA1759) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:500 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 3. Flow Cytometry analysis of HL-60 cells using anti-CACYBP antibody (PA1759).
Overlay histogram showing HL-60 cells stained with PA1759 (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-CACYBP Antibody (PA1759, 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 Cacybp (Source: Uniprot.org, NCBI)

Alternative Names

CacyBP; calcyclin binding protein; GIG5; hCacyBP; MGC87971; RP1-102G20.6; S100A6-binding protein; S100A6BPgrowth-inhibiting gene 5 protein; Siah-interacting protein (SIP); Siah-interacting protein; SIPcalcyclin-binding protein Cacybp|SIP|calcyclin binding protein|calcyclin-binding protein|siah-interacting protein

For more info on Cacybp, check out the Cacybp 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 Cacybp: 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].

Take me to the Cacybp Infographic

Here are featured tools and databases that you might find useful.

• Boster's Pathways Library
• Protein Databases
• Bioscience Research Protocol Resources
• Data Processing & Analysis Software
• Photo Editing Software
• Scientific Literature Resources
• Research Paper Management Tools
• Molecular Biology Software
• Primer Design Tools
• Bioinformatics Tools
• Phylogenetic Tree Analysis