Capicua Antibody - BSA Free

Knockout Validated: Capicua Antibody [NB110-59906] - Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and Capicua knockout (KO) HeLa cell line. PVDF membrane was probed with 1:1000 of Rabbit Anti-Human Capicua Polyclonal Antibody (Catalog# NB110-59906) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog# HAF008). Specific band was detected for Capicua at approximately 250 kDa in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. This experiment was conducted under reducing conditions.

Immunocytochemistry/Immunofluorescence: Capicua Antibody [NB110-59906] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with anti-Capicua at 5 ug/ml overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse DyLight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Western Blot: Capicua Antibody [NB110-59906] - WB detection of Human Capicua /CIC in whole cell lysates of HeLa (5, 15 and 50 ug ) and 293T (T, 50 ug) cells using affinity purified rabbit Capicua antibody NB110-59906 used at 1 ug/ml concentration.

Immunoprecipitation: Capicua Antibody [NB110-59906] - Detection of Human Capicua/CIC in 1 mg of whole cell lysate from HeLa cells (20% of IP loaded) using affinity purified rabbit polyclonal Capicua antibody (NB110-59906) at a concentration of 3 ug/mg lysate. Capicua/CIC was immunoprecipitated by NB110-59905 also, which recognizes an upstream epitope. Detection: Chemiluminescence with exposure times of 3 minutes.

Product Discontinued

Capicua Antibody - BSA Free Summary

• Immunogen: The epitope recognized by this antibody maps to a C-terminal region within residue 1500-1608 of human Capicua. [NP_055940.3]
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: CIC
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 1:50
  • Immunoprecipitation 2-5 ug/mg lysate
  • Knockout Validated
  • Western Blot 1:1000-1:2500
• Application Notes: In Western blot, a band is observed at ~250 kDa. CIC (~164 kDa) interacts with Ataxin 1 (~87 kDa) which may explain the band observed at 250 kDa (complex). In ICC/IF, nuclear staining was observed in Neuro2A cells.
• Theoretical MW: 250 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  HeLa Whole Cell Lysate

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for Capicua Antibody - BSA Free

• capicua homolog (Drosophila)
• KIAA0306capicua (Drosophila) homolog
• protein capicua homolog

Background

Capicua is a transcriptional repressor protein belonging to HMG-box proteins which has been suggested to play a key role in CNS development and is expressed in foetal brain as well as medulloblastomas. Capicua protein expression levels are subject to posttranscriptional regulation by signaling downstream of two RTKs - EGFR and Torso (that propagate their signals through Ras-Raf-MAPK signaling cascade) and MAPK-mediated phosphorylation promotes capicua degradation. Capicua has been shown to act as a transcriptional repressor through interaction with a co-repressor Groucho. Moreover, Bantam which is a Hippo pathway player has recently been shown to acts in a feedback loop to limit Capicua expression for regulating EGFR signaling. Capicua is well conserved in mammals, and mediates RTK-dependent responses that are linked to cell proliferation, cell differentiation and cancer. Capicua can interact with ATXN1 as well as ATXN1L, and has been shown to be involved in Spinocerebellar Ataxia Type 1 (SCA-1).

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Capicua Antibody (NB110-59906)(2)

Publications using NB110-59906

• Ni C, Jiang W, Wang Z et al. LncRNA-AC006129.1 reactivates a SOCS3-mediated anti-inflammatory response through DNA methylation-mediated CIC downregulation in schizophrenia Molecular Psychiatry 2020-02-03 [PMID: 32015466] (Chemotaxis, EMSA, WB, Mouse, Human)
• Yang R, Chen LH, Hansen LJ et al. Cic loss promotes gliomagenesis via aberrant neural stem cell proliferation and differentiation Cancer Res. 2017-09-22 [PMID: 28939681] (IP, Human)

Bioinformatics

• Gene Symbol: CIC
• Entrez:
  • 23152(Human)
  • 71722(Mouse)
  • 308435(Rat)
• Uniprot:
  • Q96RK0(Human)