Immunofluorescence analysis of TID1 was performed using 70% confluent log phase HepG2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with TID1 (RS13) Mouse Monoclonal Antibody (MA512267) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Mouse, Not Applicable|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||Recombinant hTid-1 protein|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunoprecipitation (IP)||2 µg/ml|
|Western Blot (WB)||1-3 µg/ml|
* 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.
|Western Blot (WB)||See 2 publications below|
MA5-12267 targets TID-1 in IP, IP, and WB applications and shows reactivity with Human, mouse, and Rat samples.
The MA5-12267 immunogen is recombinant hTid-1 protein.
TID1 is a human homolog of the Drosophila tumor suppressor lethal (2) tumerous imaginal discs, l(2)tid and encodes two mitochondrial matrix localized splice variants of human Tid-1 designated hTid-1S and hTid-1L. These proteins are the conserved members of the DnaJ family of proteins which act as cochaperons for mitochondrial Hsp70. They contain a conserved tetrahedrical J domain which binds to Hsp70 chaperones and activates their ATPase activity. Expression of hTid-1L increases apoptosis induced by DNA damaging agents as mitomycin-C and TNF-alpha. A J-domain mutant of hTid-1L can dominantly suppress apoptosis and in sharp contrast the J-domain mutant of hTid-1S increases apoptosis. Expression of hTid-1S and hTid-1L affects cytochrome c release from the mitochondria and caspase 3 activation, while activation of caspase 8 is unaffected. It is strongly suggested that these two splice variants exert their anti- and pro- apoptotic effects through discrete substrates and actiuvities. Hence the relative abundance of these proteins or their substrates may allow the mitochondria to dampen or enhance the apoptotic signals.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
p53¿ is a transcriptionally inactive p53 isoform able to reprogram cells toward a metastatic-like state.
MA5-12267 was used in western blot to study reprogramming of cells toward a metastatic-like state by p53 psi, a transcriptionally inactive p53 isoform
|Senturk S,Yao Z,Camiolo M,Stiles B,Rathod T,Walsh AM,Nemajerova A,Lazzara MJ,Altorki NK,Krainer A,Moll UM,Lowe SW,Cartegni L,Sordella R||Proceedings of the National Academy of Sciences of the United States of America (111:E3287)||2014|
Functional genetic screen for genes involved in senescence: role of Tid1, a homologue of the Drosophila tumor suppressor l(2)tid, in senescence and cell survival.
MA5-12267 was used in western blot to perform a functional genetic screen for genes involved in senescence
|Tarunina M,Alger L,Chu G,Munger K,Gudkov A,Jat PS||Molecular and cellular biology (24:10792)||2004|