AM04
Anti-Bak (Ab-2) Mouse mAb (TC-102)
Synonym(s):
Anti-Bcl-2 Antagonist Killer
antibody form
purified antibody
clone
TC-102, monoclonal
form
liquid
contains
≤0.1% sodium azide as preservative
species reactivity
mouse, human
isotype
IgG2b
General description
Anti-Bak (Ab-2), mouse monoclonal, clone TC-102, recognizes the ~30 kDa Bak protein in A431, HeLa, MCF-7, and SKBR3 cells. It is validated for Western blotting and immunofluorescence.
Bak (for bcl-2 homologous antagonist/killer) is a member of the bcl-2 family of proteins. It was independently identified by three different groups using either the two hybrid system for identifying adenovirus E1B 19K interacting proteins or by PCR cloning using degenerate BH1 and BH2 domain primers derived from the human bcl-2 gene. Bak shows a 53% homology with bcl-2 in the BH1 and BH2 domains but an overall homology of 28% with bcl-2. The presence of a hydrophobic tail on the protein suggests that, like bcl-2, it is membrane localized. The bak protein is translated from a 2.2 kb mRNA to yield a 211-216 amino acid protein with a predicted molecular weight of 23.4 kDa, but which migrates by gel electrophoresis as an approximately 30 kDa protein under denaturing conditions. Bak is expressed widely in many tissues including heart, thymus, and colon. The bak gene (bak-1) has been mapped to chromosome 6 and spans approximately 6 kb of DNA3. Southern analysis has indicated the presence of two additional bak genes: bak-2 has 97% homology with bak-1 and maps to chromosome 20, and bak-3, a pseudogene, which maps to chromosome 11. Functionally, bak is similar to bax in that it induces apoptosis. The protein pairs with bcl-xL but unlike its pairing partner has a much more extensive tissue distribution, being present in most tissues tested. While conflicting reports exist for the heterodimerization of bak with other members of the bcl-2 family of proteins, it is clear that bak does partner with the E1B 19K protein.
Purified mouse monoclonal antibody generated by immunizing mice with the specified immunogen and fusing splenocytes with Sp2/0 mouse myeloma cells. Recognizes the ~30 kDa (apparent MW) Bak protein.
Recognizes the ~30 kDa (apparent MW) Bak protein in A431, HeLa, MCF-7 or SKBR3 cells.
Immunogen
Epitope: within the N-terminal region
human recombinant Bak protein from which the C-terminal and transmembrane domains have been deleted
Application
Immunoblotting (5 μg/ml)
Immunofluorescence (1 μg/ml)
Immunofluorescence (1 μg/ml)
Physical form
In 0.05 M sodium phosphate buffer, 0.2% gelatin, pH 7.5.
Analysis Note
Negative Control
SKBR3 cells
SKBR3 cells
Positive Control
A431 cells, HeLa cells
A431 cells, HeLa cells
Other Notes
Antibody should be titrated for optimal results in individual systems for each application.
Regulatory Information
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S N Farrow et al.
Nature, 374(6524), 731-733 (1995-04-20)
A number of DNA viruses carry apoptosis-inhibiting genes which enable the virus to escape from the host response. The adenovirus E1B 19K protein can inhibit apoptosis induced by E1A, tumour-necrosis factor-alpha, FAS antigen and nerve growth factor deprivation. The molecular
M C Kiefer et al.
Nature, 374(6524), 736-739 (1995-04-20)
Members of the Bcl-2 family of proteins are characterized by their ability to modulate cell death. Bcl-2 and some of its homologues inhibit apoptosis, whereas other family members, such as Bax, will accelerate apoptosis under certain conditions. Here we describe
S J Korsmeyer et al.
Seminars in cancer biology, 4(6), 327-332 (1993-12-01)
The maintenance of homeostasis in normal tissues reflects a balance between cell proliferation and cell death. The importance of both positive and negative regulators of cell growth has been well documented in neoplasia. Bcl-2 argues for the existence of a
T Chittenden et al.
Nature, 374(6524), 733-736 (1995-04-20)
Cells are eliminated in a variety of physiological settings by apoptosis, a genetically encoded process of cellular suicide. Apoptosis comprises an intrinsic cellular defence against tumorigenesis, which, when suppressed, may contribute to the development of malignancies. The bcl-2 oncogene, which
Bcl-2 and the regulation of programmed cell death.
J C Reed
The Journal of cell biology, 124(1-2), 1-6 (1994-01-01)
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