The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/2000. Detects a band of approximately 43 kDa (predicted molecular weight: 43 kDa).
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Involved in alternative pre-RNA splicing and regulation of mRNA translation by binding to AU-rich elements (AREs) located in mRNA 3' untranslated regions (3' UTRs). Possesses nucleolytic activity against cytotoxic lymphocyte target cells. May be involved in apoptosis.
Contains 3 RRM (RNA recognition motif) domains.
Cytoplasmic granule. Nucleus. Accumulates in cytoplasmic stress granules (SG) following cellular damage.
TIA 1 cytotoxic granule associated RNA binding protein antibody
TIA1 cytotoxic granule associated RNA binding protein antibody
TIA1 cytotoxic granule associated RNA binding protein like 1 antibody
TIA1 protein antibody
Western blot - Anti-TIA1 antibody [EPR9304] (HRP) (ab195526)
All lanes : Anti-TIA1 antibody [EPR9304] (HRP) (ab195526) at 1/2000 dilution
Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate Lane 2 : MOLT4 (Human acute lymphoblastic leukemia cell line) Whole Cell Lysate Lane 3 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 43 kDa Observed band size : 43 kDa
Exposure time : 20 minutes
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab195526 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.
IHC image of TIA1 staining in a section of formalin-fixed paraffin-embedded normal human spleen tissue*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins, and incubated overnight at +4°C with ab195526 at 1/100 dilution. DAB was used as the chromogen (ab103723), diluted 1/100 and incubated for 10min at room temperature. The section was counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre