Product Name
TOM70 Recombinant Rabbit Monoclonal Antibody [PSH13-00] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within human TOM70 aa 86-608 / 608.
Species Reactivity
Human, Mouse, Rat, Green monkey
Validated Applications
WB, IHC-P, IF-Cell, IP, FC(Intra)
Target Molecular Weight
Predicted band size: 67 kDa
Positive Control
MCF7 cell lysate, HeLa cell lysate, HepG2 cell lysate, Jurkat cell lysate, MDA-MB-231 cell lysate, NIH/3T3 cell lysate, 4T1 cell lysate, C6 cell lysate, PC-12 cell lysate, COS-1 cell lysate, human colon tissue, mouse colon tissue, rat colon tissue, HeLa, NIH/3T3, C6.
Storage Instructions
Store at 2-8℃. Avoid freeze.
Purification Method
Protein A affinity purified.
Function
TOM70 (Translocase of Outer Mitochondrial Membrane 70), encoded by the TOMM70 gene, is a crucial component of the translocase of the outer mitochondrial membrane (TOM complex). The TOM complex is responsible for the recognition, unfolding, and translocation of mitochondrial preproteins synthesized in the cytosol into the mitochondria. TOM70 functions as a receptor that recognizes and mediates the import of these preproteins in a chaperone-dependent manner, playing a vital role in mitochondrial biogenesis and function.
Beyond its role in protein import, TOM70 is involved in cellular antiviral responses. Upon Sendai virus infection, TOM70 mediates the activation of TBK1 and IRF3 through MAVS, promoting host antiviral responses. It also recruits the HSP90AA1:IRF3:BAX complex to mitochondria, inducing apoptosis in response to viral infection. Additionally, TOM70 has been implicated in Parkinson's disease, as it participates in recruiting PINK1 and Parkin to depolarized mitochondria, marking them for degradation. Notably, TOM70 is targeted by the SARS-CoV-2 protein Orf9b, which suppresses interferon responses, highlighting its role in viral immune evasion.
TOM70 is widely expressed in various tissues and is essential for maintaining mitochondrial homeostasis and cellular defense mechanisms. Its multifunctional roles in mitochondrial protein import, antiviral defense, and mitochondrial quality control make it a critical protein in cellular physiology and disease.
Background References
1. Chacinska A, et al. Importing mitochondrial proteins: machineries and mechanisms. Cell. 2009, Aug.
2. Young JC, et al. Molecular chaperones Hsp90 and Hsp70 deliver preproteins to the mitochondrial import receptor Tom70. Cell. 2003 Jan.
Subcellular Location
Mitochondrion outer membrane.
Synonyms
FLJ9047 antibody
Mitochondrial import receptor subunit TOM70 antibody
Mitochondrial precursor proteins import receptor antibody
TOM70 antibody
TOM70_HUMAN antibody
Tomm70a antibody
Translocase of outer membrane 70 kDa subunit antibody
Translocase of outer membrane TOM70 antibody
Translocase of outer mitochondrial membrane 70 homolog A antibody
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This data was developed using HA723505, the same antibody clone in a different buffer formulation.
Western blot analysis of TOM70 on different lysates with Rabbit anti-TOM70 antibody (HA723505) at 1/5,000 dilution.
Lane 1: MCF7 cell lysate
Lane 2: HeLa cell lysate
Lane 3: HepG2 cell lysate
Lane 4: Jurkat cell lysate
Lane 5: MDA-MB-231 cell lysate
Lane 6: NIH/3T3 cell lysate
Lane 7: 4T1 cell lysate
Lane 8: C6 cell lysate
Lane 9: PC-12 cell lysate
Lane 10: COS-1 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 67 kDa
Observed band size: 67 kDa
Exposure time: 35 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723505) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
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This data was developed using HA723505, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-TOM70 antibody (HA723505) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723505) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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This data was developed using HA723505, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-TOM70 antibody (HA723505) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723505) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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This data was developed using HA723505, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-TOM70 antibody (HA723505) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723505) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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This data was developed using HA723505, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of HeLa cells labeling TOM70 with Rabbit anti-TOM70 antibody (HA723505) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-TOM70 antibody (HA723505) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
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This data was developed using HA723505, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of NIH/3T3 cells labeling TOM70 with Rabbit anti-TOM70 antibody (HA723505) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-TOM70 antibody (HA723505) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
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This data was developed using HA723505, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of C6 cells labeling TOM70 with Rabbit anti-TOM70 antibody (HA723505) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-TOM70 antibody (HA723505) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
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This data was developed using HA723505, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of HeLa cells labeling TOM70.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA723505, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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This data was developed using HA723505, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of NIH/3T3 cells labeling TOM70.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA723505, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
-
This data was developed using HA723505, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of C6 cells labeling TOM70.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA723505, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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This data was developed using HA723505, the same antibody clone in a different buffer formulation.
TOM70 was immunoprecipitated from 0.2 mg MCF7 cell lysate with HA723505 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723505 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: MCF7 cell lysate (input)
Lane 2: HA723505 IP in MCF7 cell lysate
Lane 3: Rabbit IgG instead of HA723505 in MCF7 cell lysate
Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 2 seconds; ECL: K1801
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