Anti-TOMM20 antibody [4F3] - BSA and Azide free

• WB

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Western blot - Anti-TOMM20 antibody [4F3] - BSA and Azide free (AB56783)

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-TOMM20 antibody [4F3] - BSA and Azide free (ab56783)

Lane 1:

HeLa cell lysate

Lane 2:

PC-12 cell lysate

Lane 3:

NIH/3T3 cell lysate

Predicted band size: 16 kDa

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• ICC/IF

AbReview19070****

Immunocytochemistry/ Immunofluorescence - Anti-TOMM20 antibody [4F3] - BSA and Azide free (AB56783)

ab56783 staining TOMM20 in HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with PBS + 0.1% Triton X-100 and blocked with PBS+ 0.5% BSA + 0.2% fish skin gelatin for 1 hour at 25°C. Samples were incubated with primary antibody (1/2000 in PBS + 1% BSA) for 24 hours at 4°C. An Alexa Fluor^® 488-conjugated goat anti-mouse IgG polyclonal was used as the secondary antibody (1/500).

This image was generated using the ascites version of the product.

This image is courtesy of an anonymous Abreview.

• ICC/IF

AbReview24004****

Immunocytochemistry/ Immunofluorescence - Anti-TOMM20 antibody [4F3] - BSA and Azide free (AB56783)

ab56783 staining TOMM20 in human keratinocytes by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with acetone and blocked with 1% BSA for 1 hour at 24°C. Samples were incubated with primary antibody (1/2000 in PBS + 1% BSA) for 24 hours at 4°C. An Alexa Fluor^® 488-conjugated goat anti-mouse IgG polyclonal was used as the secondary antibody (1/5000).

This image was generated using the ascites version of the product.

This image is courtesy of an anonymous Abreview.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-TOMM20 antibody [4F3] - BSA and Azide free (AB56783)

Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling TOMM20 with ab56783 at 10 μg/ml. Cells are fixed with 4% PFA and permeabilized on ice in PBS 0.1% Triton. Samples were incubated with primary antibody at 4^oC overnight and fluorscein- conjugated secondary antibody at 4^oC for 1 hour.

This image was generated using the ascites version of the product.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TOMM20 antibody [4F3] - BSA and Azide free (AB56783)

Paraffin embedded human small intestine tissue stained for TOMM20 using ab56783 at 3 μg/ml in immunohistochemical analysis.

• Flow Cyt

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Flow Cytometry - Anti-TOMM20 antibody [4F3] - BSA and Azide free (AB56783)

Overlay histogram showing HeLa cells stained with ab56783 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab56783, 1 μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was a mix of mouse IgG1 [ICIGG1], (ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4%PFA/permeabilized in 0.1% PBS-Tween used under the same conditions.

This image was generated using the ascites version of the product.

• IP

Unknown

Immunoprecipitation - Anti-TOMM20 antibody [4F3] - BSA and Azide free (AB56783)

TOMM20 was immunoprecipitated using 0.5 mg HepG2 whole cell extract, 5 μg of Mouse monoclonal to TOMM20 (ab56783) and 50 μl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10 min, HepG2 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10 min under agitation.
Proteins were eluted by addition of 40 μl SDS loading buffer and incubated for 10 min at 70°C; 10 μl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab56783.
Secondary : Protein G-HRP at 1/500 dilution.
Band : 14kDa : TOMM20. Non specific - 25kDa : We are unsure as to the identity of this extra band.

This image was generated using the ascites version of the product.

All lanes:

Immunoprecipitation - Anti-TOMM20 antibody [4F3] - BSA and Azide free (ab56783)

Predicted band size: 16 kDa

false