Western blot analysis of extracts from various cell lines using FANCA (D1L2Z) Rabbit mAb (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). As expected, GM06914 cells are negative for FANCA.
Western blot analysis of extracts from various cell types using FANCB (D9W6S) Rabbit mAb.
Western blot analysis of extracts fro HeLa, U-2 OS, and GM16633 (PD20, FANCD2-negative) cells using FANCD2 (D5L5X) Rabbit mAb (upper) or α-Actinin (D6S6) Rabbit mAb #6487 (lower).
Western blot analysis of extracts from various cell lines using BRCA2 (D9S6V) Rabbit mAb (upper) or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
Western blot analysis of extracts from SH-SY5Y, Saos-2 and Jurkat cells using BACH1/BRIP1 Antibody.
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Immunoprecipitation of FANCA from 293T cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or FANCA (D1L2Z) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot was performed using FANCA (D1L2Z) Rabbit mAb.
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human FANCB (FANCB-Myc/DDK; +) protein, using FANCB (D9W6S) Rabbit mAb.
Western blot analysis of extracts from HeLa cells, untreated or treated with mitomycin C (2 μg/mL, 24 hr), using FANCD2 (D5L5X) Rabbit mAb. DNA damage caused by mitomycin C induces monoubiquitination of FANCD2, altering its electrophoretic mobility and increasing its apparent molecular weight (4).
Western blot analysis of DLD1 cells, either wild type (WT) or BRCA2 knockout (-/-), using BRCA2 (D9S6V) Rabbit mAb (upper) or α-Actinin (D6F6) XP® Rabbit mAb (lower). DLD1 (WT) and DLD1 (-/-) cells were kindly provided by Dr. Judit Jimenez, Yale University, New Haven, CT.
Immunoprecipitation of FANCD2 from HeLa cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb XP® Isotype Control #3900, and lane 3 is FANCD2 (D5L5X) Rabbit mAb. Western blot analysis was performed using FANCD2 (D5L5X) Rabbit mAb.
|FANCA (D1L2Z) Rabbit mAb 14657||20 µl||
|FANCB (D9W6S) Rabbit mAb 14243||20 µl||
||H M R||90||Rabbit IgG|
|FANCD2 (D5L5X) Rabbit mAb 16323||20 µl||
||H||155, 162||Rabbit IgG|
|BRCA2 (D9S6V) Rabbit mAb 10741||20 µl||
|BACH1/BRIP1 Antibody 4578||20 µl||
|Anti-rabbit IgG, HRP-linked Antibody 7074||100 µl||
The Fanconi Anemia Antibody Sampler Kit provides an economical means of detecting members of the Fanconi Anemia signaling pathway. The kit includes enough antibody to perform two western blot experiments with each primary antibody.
Each antibody in the Fanconi Anemia Antibody Sampler kit detects endogenous levels of its target protein.
Monoclonal antibodies are produced by immunizing rabbits with synthetic peptides corresponding to residues surrounding Ala514 of human FANCA, Pro280 of human FANCB, Gly995 of human FANCD2, and a recombinant protein specific to the carboxy terminus of human BRCA2. Polyclonal BACH1/BRIP1 Antibody is produced by immunizing rabbits with a synthetic peptide corresponding to amino acids near the carboxy terminus of human BACH1. Polyclonal antibodies are purified by Protein A and peptide affinity chromatography.
Fanconi anemia (FA) is an autosomal recessive genetic disorder resulting in symptoms that include chromosomal breakage, bone marrow failure, hypersensitivity to DNA cross-linking agents (such as mitomycin C), and a predisposition to cancer (1). In response to DNA damage, the FA nuclear complex (FANCA, FANCB, FANCC, FANCE, FANCF, FANCG, FANCM) induces mono-ubiquitination of FANCD2 and FANCI (2).
Monoubiquitination of FANCD2 induces localization of FANCD2 to sites of DNA damage, where it interacts with BRCA1 (4). FANCJ/BRIP1, FANCD1/BRCA2, and FANCN/PALB2 are also recruited to sites of DNA damage. FA signaling is important in maintenance of chromosome stability and control of mitosis (3).
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