Anti-FLI1 antibody [EPR4646]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FLI1 antibody [EPR4646] (AB133485)

Immunohistochemical analysis of formalin fixed paraffin embedded human lung labelling FLI1 with ab133485 at a concentration of 0.01 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH 8.5 for 32mins.

ab133485 Anti-FLI1 antibody [EPR4646] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FLI1 antibody [EPR4646] (AB133485)

Immunohistochemical analysis of paraffin-embedded Human Ewing sarcoma tissue labelled with unpurified ab133485 at 1/250 dilution.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FLI1 antibody [EPR4646] (AB133485)

Immunohistochemical staining of paraffin embedded human lung with unpurified ab133485 at a working dilution of 1 in 140. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FLI1 antibody [EPR4646] (AB133485)

Immunohistochemical staining of paraffin embedded human lung with purified ab133485 at a working dilution of 1 in 700. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-FLI1 antibody [EPR4646] (AB133485)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ A-673 (human muscle Ewings sarcoma cell line) cells and 5 µg of ab133485 [EPR4646]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-FLI1 antibody [EPR4646] (AB133485)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ A-673 (human muscle Ewings sarcoma cell line) cells and 5 µg of ab133485 [EPR4646]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-FLI1 antibody [EPR4646] (AB133485)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ A-673 (human muscle Ewings sarcoma cell line) cells and 5 µg of ab133485 [EPR4646]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• WB

Lab

Western blot - Anti-FLI1 antibody [EPR4646] (AB133485)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-FLI1 antibody [EPR4646] (ab133485) at 1/1000 dilution

All lanes:

Jurkat cell lysate at 10 µg

Secondary

All lanes:

HRP goat anti-rabbit at 1/1000 dilution

Predicted band size: 51 kDa

Observed band size: 51 kDa

false

• WB

Unknown

Western blot - Anti-FLI1 antibody [EPR4646] (AB133485)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-FLI1 antibody [EPR4646] (ab133485) at 1/200 dilution

All lanes:

Jurkat cell lysate at 10 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 51 kDa

Observed band size: 51 kDa

false

• WB

Unknown

Western blot - Anti-FLI1 antibody [EPR4646] (AB133485)

All lanes:

Western blot - Anti-FLI1 antibody [EPR4646] (ab133485) at 1/1000 dilution

Lane 1:

U937 cell lysate at 10 µg

Lane 2:

Jurkat cell lysate at 10 µg

Lane 3:

Raw264.7 cell lysate at 10 µg

Lane 4:

HL-60 cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 51 kDa

false

• WB

Unknown

Western blot - Anti-FLI1 antibody [EPR4646] (AB133485)

All lanes:

Western blot - Anti-FLI1 antibody [EPR4646] (ab133485) at 1/1000 dilution

Lane 1:

FLI1 recombinant protein at 0.01 µg

Lane 2:

ERGrecombinant protein at 0.01 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 51 kDa

false