Anti-TFEB antibody [EPR22940-151] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-TFEB antibody [EPR22940-151] - BSA and Azide free (AB270614)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab270604).

Flow cytometry overlay histogram showing left Raji positive cells and right negative MCF7 stained with ab270604 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab270604) (1x 106 in 100μl at 0.2μg/ml (1/10350)) for 30min at 22°C.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C

Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-TFEB antibody [EPR22940-151] - BSA and Azide free (AB270614)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Raji (Human Burkitt's lymphoma B lymphocyte) cells labeling TFEB with ab270604 at 1/500 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^®488, ab150077) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab270604).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TFEB antibody [EPR22940-151] - BSA and Azide free (AB270614)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling TFEB with ab270604 at 1/500 followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in human colon tissue is observed (PMID : 30519051). The section was incubated with ab270604 for 20 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab270604).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-TFEB antibody [EPR22940-151] - BSA and Azide free (AB270614)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized IM-9 cells labeling TFEB with ab270604 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in IM-9 cells. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab270604).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TFEB antibody [EPR22940-151] - BSA and Azide free (AB270614)

Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling TFEB with ab270604 at 1/500 followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in human lung cancer tissue is observed (PMID : 26264650). The section was incubated with ab270604 for 20 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab270604).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-TFEB antibody [EPR22940-151] - BSA and Azide free (AB270614)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized IM-9 (Human multiple myeloma B Lymphoblast) cells labeling TFEB with ab270604 at 1/500 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^®488, ab150077) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab270604).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-TFEB antibody [EPR22940-151] - BSA and Azide free (AB270614)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Raji cells labeling TFEB with ab270604 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in Raji cells. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab270604).

• IP

Unknown

Immunoprecipitation - Anti-TFEB antibody [EPR22940-151] - BSA and Azide free (AB270614)

TFEB was immunoprecipitated from 0.35 mg Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate with ab270604 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab270604 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/1000 dilution.

Lane 1 : Raji whole cell lysate.

Lane 2 : ab270604 IP in Raji whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab270604 in Raji whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 3 secs.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab270604).

All lanes:

Immunoprecipitation - Anti-TFEB antibody [EPR22940-151] (<a href='/en-us/products/primary-antibodies/tfeb-antibody-epr22940-151-ab270604'>ab270604</a>)

Predicted band size: 53 kDa

Observed band size: 66 kDa

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• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-TFEB antibody [EPR22940-151] - BSA and Azide free (AB270614)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (,ab270604).

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 Raji (human Burkitt's lymphoma B lymphocyte) cells and 5 µg of ab270604 [EPR22940-151]. 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-TFEB antibody [EPR22940-151] - BSA and Azide free (AB270614)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (,ab270604).

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 Raji (human Burkitt's lymphoma B lymphocyte) cells and 5 µg of ab270604 [EPR22940-151]. 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-TFEB antibody [EPR22940-151] - BSA and Azide free (AB270614)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (,ab270604).

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 Raji (human Burkitt's lymphoma B lymphocyte) cells and 5 µg of ab270604 [EPR22940-151]. 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.