ANTIBODIES TO CONDENSIN COMPLEX PROTEINS
Precise duplication and segregation of chromosomes is crucial to cellular
growth and organismal development. The evolutionary conservation of proteins
and protein complexes involved in this process attests to its importance.
Cohesin and condensin are two classes of multisubunit complexes required for
this critical process. Both human complexes contain a specific heterodimer of
SMC (structural maintenance of chromosomes) proteins that constitute the core
and additional non-SMC subunits that complete the complex.
The condensin complex is a heteropentameric complex that participates in the
condensation of chromosomes during cell division by facilitating positive DNA
supercoiling and knotting by topoisomerase I and II. There are two condensin
complexes which are defined by their non-SMC regulatory subunits. Condensin I
is comprised of SMC2, SMC4, CAP-D2, CAP-G, and CAP-H, while condensin II is
comprised of SMC2, SMC4, CAP-D3, CAP-G2, and CAP-H2. A basic understanding of
the specific roles of some of these subunits has been demonstrated. For
example, CAP-D2 may be important for targeting condensin to mitotic chromosomes
and metaphase alignment of sister chromatids, CAP-D3 may participate in
centromere resolution, and CAPH may have an important role in condensin’s
ability to maintain structural integrity of centromeric heterochromatin during
mitosis. There is evidence that the condensins also participate in non-mitotic
functions such as transcription and replication. Research that further explores
the contributions of the subunits of condensin will lead to a better
understanding of how condensins and cohesins participate in the maintenance of
chromosomes during cell division and their importance to non-mitotic cellular
functions.
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Detection of Human CAP-D2 by
Western Blot and Immunoprecipitation.
Samples: Whole cell lysate from
HeLa (5, 15 and 50 mcg for Western Blot; 1 mg for
Immunoprecipitation, 20% of IP loaded) and 293T
(50 mcg for Western Blot) cells.
Antibodies: Affinity purified rabbit anti-CAP-D2
antibody A300-601A used
for Western Blot
at 0.04 mcg/ml (A) and 1 mcg/ml (B) and for
IP at 3 mcg/mg lysate (B). CAP-D2 was also
immunoprecipitated using a second rabbit
anti-CAP-D2 antibody, which binds an upstream
epitope. For detecting immunoprecipitated CAP-D2
ReliaBLOT® Reagents and Procedures
(Cat. No. WB120) were used.
Detection: Chemiluminescence with exposure
times of 30 seconds (A and B).
ReliaBLOT is a registered trademark of
Bethyl Laboratories, Inc. |
Detection of Human Aurora B by
Immunocytochemistry.
Sample: NBF-fixed HeLa Cells.
Antibody: Affinity purified rabbit
anti-Aurora B (Cat. No. IHC-00003)
used at a dilution of 1:100.
Detection: Goat anti-rabbit IgG
(heavy & light) FITC (Cat. No.
A120-101F)
used at a dilution of 1:100. |
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