Anti-T-bet / Tbx21 antibody [39D, 3-9D]

• ICC

Supplier Data

Immunocytochemistry - Anti-T-bet / Tbx21 antibody [39D, 3-9D] (AB91103)

Immunocytochemistry analysis of T-bet/ Tbx21 was performed using 70% confluent log phase NK-92 cells. The cells were fixed with 4% paraformaldehyde for 10 Minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with ab91103 at 5 μg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 at a dilution of 1/2000 for 45 minutes at room temperature (Panel a : green). Nuclei (Panel b : blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI. F-actin (Panel c : red) was stained with Rhodamine Phalloidin. Panel d represents the merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

• ChIP

Supplier Data

ChIP - Anti-T-bet / Tbx21 antibody [39D, 3-9D] (AB91103)

Chromatin Immunoprecipitation (ChIP) assay of endogenous T-bet protein using Anti-T-bet Antibody : ChIP was performed using ab91103 at 5 μg on sheared chromatin from NK-92 cells using the MAGnify ChIP System kit. Normal Mouse IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to IFNG promoter, IKZF3 and CD19 transcriptional start site and SAT2 satellite repeats. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

• WB

Supplier Data

Western blot - Anti-T-bet / Tbx21 antibody [39D, 3-9D] (AB91103)

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 re-activation with PMA and Ionomycin.

All lanes:

Western blot - Anti-T-bet / Tbx21 antibody [39D, 3-9D] (ab91103) at 2 µg/mL

Lane 1:

Lysates from control cells

Lane 2:

Lysates from PMA and Ionomycin-re-activated cells

Secondary

All lanes:

HRP anti-mouse IgG

Predicted band size: 58 kDa

false

• WB

Supplier Data

Western blot - Anti-T-bet / Tbx21 antibody [39D, 3-9D] (AB91103)

All lanes:

Western blot - Anti-T-bet / Tbx21 antibody [39D, 3-9D] (ab91103) at 2 µg/mL

Lane 1:

NK-92 cell lysates at 30 µg

Lane 2:

K-562 cell lysates at 30 µg

Secondary

All lanes:

Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant, HRP at 1/4000 dilution

Predicted band size: 58 kDa

false