The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml.
Use a concentration of 2 µg/ml. Predicted molecular weight: 58 kDa.
Use at an assay dependent concentration.
Use 2µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Transcription factor that controls the expression of the TH1 cytokine, interferon-gamma. Initiates TH1 lineage development from naive TH precursor cells both by activating TH1 genetic programs and by repressing the opposing TH2 programs.
Genetic variations in TBX21 are associated with susceptibility to asthma with nasal polyps and aspirin intolerance (ANPAI) [MIM:208550]. A condition consisting of asthma, aspirin sensitivity and nasal polyposis. Nasal polyposis is due to chronic inflammation of the paranasal sinus mucosa, leading to protrusion of edematous polyps into the nasal cavities.
Western blot - T-bet / Tbx21 antibody [4B10] (ab91109)
All lanes : Anti-T-bet / Tbx21 antibody [4B10] (ab91109) at 2 µg/ml
Lane 1 : Lysates of CD4+ T cells (control) Lane 2 : Lysates of PMA and Ionomycin reactivated CD4+ T cells
Secondary HRP anti-mouse IgG
Predicted band size : 58 kDa CD4+ T cells were sorted from Mouse spleen, activated with anti-mouse CD3 and anti-mouse CD28, followed by culture in Th1 polarizing conditions, and reactivation with PMA and Ionomycin.
IHC image of T-bet / Tbx21 staining in Human normal tonsil formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab91109, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
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.
ab91109 staining T-bet in Human acne lesion tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with 10% formaldehyde and antigen retrieval was by heat mediation perfomred in a microwave oven. Samples were incubated with primary antibody (1/200).
Overlay histogram showing Jurkat cells stained with ab91109 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab91109, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Fang J et al. Prognostic significance of tumor infiltrating immune cells in oral squamous cell carcinoma. BMC Cancer17:375 (2017).
Read more (PubMed: 28549420) »
de Souza AWS et al. M2 macrophage is the predominant phenotype in airways inflammatory lesions in patients with granulomatosis with polyangiitis. Arthritis Res Ther19:100 (2017).
Read more (PubMed: 28521792) »