Anti TMBIM1  

Immunohistochemistry of Rabbit anti-TMBIM1 antibody. Tissue: human prostate. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: anti-TMBIM1 antibody at 10 μg/mL for 1 h at RT. Secondary antibody: Peroxidase rabbit secondary antibody at 1:10,000 for 45 min at RT. Staining: TMBIM1 as precipitated red signal with hematoxylin purple nuclear counterstain.

Immunohistochemistry of Rabbit anti-TMBIM1 antibody. Tissue: human tonsil. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: anti-TMBIM1 antibody at 10 μg/mL for 1 h at RT. Secondary antibody: Peroxidase rabbit secondary antibody at 1:10,000 for 45 min at RT. Staining: TMBIM1 as precipitated red signal with hematoxylin purple nuclear counterstain.

Western blot using Rockland's protein A purified anti-TMBIM1 antibody shows detection of exogenous TMBIM1 in lysates from HeLa cells transfected with pcDNA3-hTMBIM1 (lane 1). No staining is observed in lysates from mock transformed HeLa cells (lane 2). To date this antibody has shown the ability to recognize over expressed TMBIM1 ~35kDa, but not endogenous protein. The membrane was probed with the primary antibody at a 1:1,000 dilution at 4° C, overnight. Personal Communication from Srinivasa Srinivasula, CCR-NCI, Bethesda, MD.

c？ (TMBIM1) PP1201 is expressed by various cell lines. Semi-quantitative RT-PCR was performed to quantify the expression level of PP1201 in different cell lines (Upper Panel). Amplifications of glyceraldehyde-3-phosphate dehydrogenase were used as loading controls. Lower panels show endogenous protein levels in corresponding cell lines and？β-Actin was used as loading control for protein. 30 μg of protein was loaded in each well.？d？Generation of stable cell lines. Using retroviral transfection (As described in “Materials and methods”) Jurkat and HT1080 cell lines were used to stably express PP1201. Endogenous expression of PP1201 was present in HT1080 and absent in Jurkat.？β-Actin was used as loading control for protein. Fig 1. PMID:？21107705

PP1201 interacts with Fas mainly through transmembrane region and localizes into intracellular compartment.？a？Coimmunoprecipitation of PP1201 with FLAG Fas. Expression plasmids for PP1201 and FLAG Fas were transfected in combination and alone in 293 cells. Cell lysates were immunoprecipitated with mouse anti-FLAG and immunoblotted with rabbit anti-PP1201. Lysates were immunoblotted with anti-FLAG, anti-PP1201 and anti？β-actin.？b？Coimmunoprecipitation of N-terminal tagged V5 truncated PP1201-TM (expressing transmembrane region, 103？311aa) and FLAG Fas. Expression plasmids of these two were transfected in 293-T cell in combination and alone. Cell lysates were immunoprecipitated with mouse anti-FLAG and immunoblotted with mouse anti V5. Lysates were immunoblotted with anti-FLAG, anti-V5 and anti？β-actin.？c？Immunostaining for PP1201 at endogenous level in HUVEC cells using GM130 as a golgi marker shows that PP1201 partially localized into golgi.？d？Transient transfection of RFP tagged PP1201 in Hela cells and staining with golgi marker GM130 shows PP1201 to be localized into golgi compartment.？e？Transient transfection of CFP (Cyan Fluorescent Protein) tagged Fas along with RFP tagged PP1201 shows colocalization of both mostly into perinuclear spaces like Golgi. Fig 3. PMID:？21107705