Anti-TEF1/TEAD-1 antibody [EPR3967(2)]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TEF1/TEAD-1 antibody [EPR3967(2)] (AB133533)

Immunohistochemistry analysis of Paraffin Embedded Human placenta tissue labelling TEF1/TEAD-1 with unpurified ab133533 at 1/100. Heat mediated antigen retrieval was performed using citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TEF1/TEAD-1 antibody [EPR3967(2)] (AB133533)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human skeletal muscle tissue sections labeling TEF1/TEAD-1 with Purified ab133533 at 1 : 1000 dilution (1.63 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, pH 9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1 : 0 dilution. PBS instead of the primary antibody was used as the negative control.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TEF1/TEAD-1 antibody [EPR3967(2)] (AB133533)

Immunohistochemistry analysis of Paraffin Embedded Human skeletal muscle tissue labelling TEF1/TEAD-1 with unpurified ab133533 at 1/100. Heat mediated antigen retrieval was performed using citrate buffer pH 6 before commencing with IHC staining protocol.

• IP

Lab

Immunoprecipitation - Anti-TEF1/TEAD-1 antibody [EPR3967(2)] (AB133533)

ab133533 (purified) at 1 : 80 dilution (2ug) immunoprecipitating TEF1/TEAD-1 in 293 (Human embryonic kidney epithelial cell) whole cell lysate.
Lane 1 (input) : 293 (Human embryonic kidney epithelial cell) whole cell lysate 10ug
Lane 2 (+) : ab133533 & 293 (Human embryonic kidney epithelial cell) whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab133533 in 293 (Human embryonic kidney epithelial cell) whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1 : 1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-TEF1/TEAD-1 antibody [EPR3967(2)] (ab133533)

Predicted band size: 48 kDa

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• ChIP-seq

Lab

ChIP-sequencing - Anti-TEF1/TEAD-1 antibody [EPR3967(2)] (AB133533)

Chromatin was prepared from HepG2 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab133533 [EPR3967(2)]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-TEF1/TEAD-1 antibody [EPR3967(2)] (AB133533)

Chromatin was prepared from HepG2 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab133533 [EPR3967(2)]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-TEF1/TEAD-1 antibody [EPR3967(2)] (AB133533)

Chromatin was prepared from HepG2 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab133533 [EPR3967(2)]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TEF1/TEAD-1 antibody [EPR3967(2)] (AB133533)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat spleen tissue sections labeling TEF1/TEAD-1 with Purified ab133533 at 1 : 1000 dilution (1.63 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, pH 9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1 : 0 dilution. PBS instead of the primary antibody was used as the negative control.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TEF1/TEAD-1 antibody [EPR3967(2)] (AB133533)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse spleen tissue sections labeling TEF1/TEAD-1 with Purified ab133533 at 1 : 1000 dilution (1.63 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, pH 9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1 : 0 dilution. PBS instead of the primary antibody was used as the negative control.

• ChIP-seq

Lab

ChIP-sequencing - Anti-TEF1/TEAD-1 antibody [EPR3967(2)] (AB133533)

Chromatin was prepared from MEF cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab133533 [EPR3967(2)]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-TEF1/TEAD-1 antibody [EPR3967(2)] (AB133533)

Chromatin was prepared from MEF cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab133533 [EPR3967(2)]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• WB

Lab

Western blot - Anti-TEF1/TEAD-1 antibody [EPR3967(2)] (AB133533)

Blocking/Diluting buffer and concentration : 5% NFDM/TBST. ab133533 recognizes human TEAD1 and human TEAD3.

All lanes:

Western blot - Anti-TEF1/TEAD-1 antibody [EPR3967(2)] (ab133533) at 1/5000 dilution

Lane 1:

His-GST-tagged human TEAD1 recombinant protein at 20 µg

Lane 2:

His-SUMO-tagged human TEAD3 recombinant protein at 20 µg

Lane 3:

His-tagged human TEAD4 recombinant protein at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 50 kDa,70 kDa

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Exposure time: 10s

• WB

Lab

Western blot - Anti-TEF1/TEAD-1 antibody [EPR3967(2)] (AB133533)

Lanes 1 - 2 : Merged signal (red and green). Green - ab133533 observed at 50 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab133533 was shown to react with TEF1/TEAD-1 in wild-type A549 cells in western blot with loss of signal observed in TEAD1 knockout sample. Wild-type and TEAD1 knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab133533 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-TEF1/TEAD-1 antibody [EPR3967(2)] (ab133533) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

TEAD1 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Predicted band size: 48 kDa

Observed band size: 50 kDa

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• WB

Lab

Western blot - Anti-TEF1/TEAD-1 antibody [EPR3967(2)] (AB133533)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-TEF1/TEAD-1 antibody [EPR3967(2)] (ab133533) at 1/2000 dilution

All lanes:

Human fetal muscle lysates at 15 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 48 kDa

Observed band size: 52 kDa

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• WB

Unknown

Western blot - Anti-TEF1/TEAD-1 antibody [EPR3967(2)] (AB133533)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-TEF1/TEAD-1 antibody [EPR3967(2)] (ab133533) at 1/10000 dilution

Lane 1:

L6 ( Rat skeletal muscle myoblast) whole cell lysates at 15 µg

Lane 2:

C2C12 ( Mouse myoblasts myoblast) whole cell lysate at 15 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 48 kDa

Observed band size: 52 kDa

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• WB

Unknown

Western blot - Anti-TEF1/TEAD-1 antibody [EPR3967(2)] (AB133533)

All lanes:

Western blot - Anti-TEF1/TEAD-1 antibody [EPR3967(2)] (ab133533) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

293T cell lysate at 10 µg

Lane 3:

Fetal muscle lysate at 10 µg

Predicted band size: 48 kDa

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• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-TEF1/TEAD-1 antibody [EPR3967(2)] (AB133533)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.