Immunofluorescent analysis of TCP1 (green) in murine cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes at room temperature and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a TCP1 Monclonal Antibody (91A) (Product # MA3-026 ) at a dilution of 1:100 and incubated overnight in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody for 45 minutes at room temperature in the dark. F-actin (red) was stained with a fluorescent phalloidin and nuclei (blue) were stained with DAPI. Images were taken at a 60X magnification.
|Tested species reactivity||Dog, Chicken, Hamster, Human, Mouse, Non-human primate, Rat, Yeast|
|Published species reactivity||Mouse|
|Host / Isotype||Rat / IgG2a|
|Immunogen||Bacterially expressed fragment of mouse TCP-1, residues 306-556.|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1/100|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1:1,000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Immunoprecipitation (IP)||See 1 publications below|
MA3-026 detects T Complex Polypeptide 1 (TCP-1) from human, canine, chicken, hamster, mouse, primate, rat and yeast samples.
MA3-026 has been successfully used in Western blot, immunofluorescence and immunoprecipitation procedures. By Western blot, this clone detects a ~57 kDa band representing TCP-1 in mouse 3T3 cells. Immunofluorescence staining of TCP-1 in mouse germ line cells with MA3-026 results in diffuse cytoplasmic staining.
The MA3-026 immunizing peptide corresponds to amino acid residues 306-556 from mouse TCP-1. The epitope for this antibody has been mapped to amino acids 465-469 representing the pentamer peptide AKLRA.
The T Complex Polypeptide 1 (TCP-1) is a ~60 kDa protein constitutively expressed in almost all eukaryotic cells, and is upregulated during spermatogenesis. It is found in the cytosol as a subunit of a hetero-oligomeric chaperone that is known to be involved in the folding of actin and tubulin. The family of proteins termed chaperonins act to recognize and stabilize polypeptide intermediates during folding, assembly and disassembly, and share many characteristics with Heat Shock Protein 70 (HSP70) including high abundance, induction by environmental stress, and ATPase activity. The chaperonin family includes the mitochondrial HSP60, Escherichia coli GroEL, the plastid Rubisco-subunit binding protein, and the archaebacterial protein TF55. The TCP-1 sequence shows nearly 40% identity to TF55, but only minimal similarity to HSP60 and GroEL.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Identification of six Tcp-1-related genes encoding divergent subunits of the TCP-1-containing chaperonin.
MA3-026 was used in immunoprecipitation to characterize different subunits of CCT chaperonin
|Kubota H,Hynes G,Carne A,Ashworth A,Willison K||Current biology : CB (4:89)||1994|