Anti-mtTFA antibody [EPR12285] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mtTFA antibody [EPR12285] - BSA and Azide free (AB240958)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human kidney tissue, labeling mtTFA using unpurified ab176558 at a 1/50 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab176558).

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-mtTFA antibody [EPR12285] - BSA and Azide free (AB240958)

Immunofluorescence staining of MCF7 cells with purified ab176558 at a working dilution of 1/250, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor^® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100.

The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab176558 was used at a dilution of 1/500 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500.

For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor^® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab176558).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-mtTFA antibody [EPR12285] - BSA and Azide free (AB240958)

Immunofluorescence analysis of MCF7 cells, labeling mtTFA using unpurified ab176558 at a 1/100 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab176558).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mtTFA antibody [EPR12285] - BSA and Azide free (AB240958)

Immunohistochemical staining of paraffin embedded human thyroid carcinoma with purified ab176558 at a working dilution of 1/300. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab176558).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mtTFA antibody [EPR12285] - BSA and Azide free (AB240958)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human prostate tissue, labeling mtTFA using unpurified ab176558 at a 1/50 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab176558).

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IP

Unknown

Immunoprecipitation - Anti-mtTFA antibody [EPR12285] - BSA and Azide free (AB240958)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab176558).
Purified ab176558 at 1 : 40 dilution immunoprecipitating mtTFA in HEK-293 whole cell lysate.
Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Lane 1:

Immunoprecipitation - Anti-mtTFA antibody [EPR12285] - Mitochondrial Marker (<a href='/en-us/products/primary-antibodies/mttfa-antibody-epr12285-mitochondrial-marker-ab176558'>ab176558</a>)

Lanes 1 - 2:

Immunoprecipitation - Anti-mtTFA antibody [EPR12285] - Mitochondrial Marker (<a href='/en-us/products/primary-antibodies/mttfa-antibody-epr12285-mitochondrial-marker-ab176558'>ab176558</a>) at 1/40 dilution

Lane 1:

HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate at 10 µg

Lane 2:

<a href='/en-us/products/primary-antibodies/mttfa-antibody-epr12285-mitochondrial-marker-ab176558'>ab176558</a> + HEK-293 whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/en-us/products/primary-antibodies/mttfa-antibody-epr12285-mitochondrial-marker-ab176558'>ab176558</a> in HEK-293 whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 29 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-mtTFA antibody [EPR12285] - BSA and Azide free (AB240958)

Western blot analysis on Immunoprecipitation pellet from either 1) 293T cell lysate, or 2) 1xPBS (negative control), showing mtTFA, using unpurified ab176558 at 1/10 dilution and HRP-conjugated anti-rabbit IgG preferentially detecting the non-reduced form of rabbit IgG.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab176558).

All lanes:

Immunoprecipitation - Anti-mtTFA antibody [EPR12285] - Mitochondrial Marker (<a href='/en-us/products/primary-antibodies/mttfa-antibody-epr12285-mitochondrial-marker-ab176558'>ab176558</a>)

Predicted band size: 29 kDa

true