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AB258635

Human RHOT1 (MIRO1) knockout HeLa cell lysate

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RHOT1 KO cell lysate available now. KO validated. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon1.

View Alternative Names

ARHT 1, FLJ11040, FLJ12633, MIRO1_HUMAN, Mitochondrial Rho 1, Mitochondrial Rho GTPase 1, Rac-GTP-binding protein-like protein, Ras homolog family member T1, Ras homolog gene family member T1, Rhot 1, hMiro-1, mitochondrial Rho (MIRO) GTPase 1

2 Images
Western blot - Human RHOT1 (MIRO1) knockout HeLa cell lysate (AB258635)
  • WB

Lab

Western blot - Human RHOT1 (MIRO1) knockout HeLa cell lysate (AB258635)

Lane 1 : Wild-type HeLa cell lysate 40 μg
Lane 2 : RHOT1 knockout HeLa cell lysate 40 μg
Lane 3 : HEK-293 cell lysate 20 μg
Lane 4 : A431 cell lysate 20 μg
False colour image of Western blot : Anti-MIRO1 antibody [CL1083] staining at 1/500 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab188029 was shown to bind specifically to MIRO1. A band was observed at 71 kDa in wild-type HeLa cell lysates with no signal observed at this size in RHOT1 knockout cell line ab265792 (knockout cell lysate ab258635). To generate this image, wild-type and RHOT1 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

All lanes:

Western blot - Anti-MIRO1 antibody [CL1083] (<a href='/en-us/products/primary-antibodies/miro1-antibody-cl1083-ab188029'>ab188029</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 40 µg

Lane 2:

RHOT1 knockout HeLa cell lysate at 40 µg

Lane 2:

Western blot - Human RHOT1 (MIRO1) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-rhot1-miro1-knockout-hela-cell-line-ab265792'>ab265792</a>)

Lane 3:

HEK-293 cell lysate at 20 µg

Lane 4:

A431 cell lysate at 20 µg

Predicted band size: 71 kDa

Observed band size: 71 kDa

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Sanger Sequencing - Human RHOT1 (MIRO1) knockout HeLa cell lysate (AB258635)
  • Sanger seq

Unknown

Sanger Sequencing - Human RHOT1 (MIRO1) knockout HeLa cell lysate (AB258635)

Homozygous : 1 bp insertion in exon1

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon1.

Disease

Adenocarcinoma

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

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What's included?

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Properties and storage information

Gene name
RHOT1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

MIRO1 also known as RhoT1 is a mitochondrial Rho GTPase with a molecular mass of approximately 69 kDa. It functions as a pivotal component in mitochondrial transport and dynamics. This protein localizes to the outer mitochondrial membrane and is highly expressed in various tissues particularly in the brain and muscles. MIRO1 facilitates the anchoring of mitochondria to microtubule motors enabling their movement within the cell which is essential for maintaining proper energy distribution.
Biological function summary

MIRO1 plays a significant role in regulating mitochondrial trafficking along the cytoskeleton. It forms part of a protein complex with TRAK1/2 kinesin and dynein motor proteins. MIRO1's presence in this complex allows it to modulate mitochondrial distribution and dynamics which are essential for neural development and function. It also influences mitochondrial shape and interaction with the endoplasmic reticulum impacting intracellular calcium homeostasis.

Pathways

MIRO1 is an integral component of pathways involving mitochondrial transport and cellular energy homeostasis. It actively participates in the regulation of calcium signaling pathways alongside related proteins like TRAK1. Additionally MIRO1's involvement in the maintenance of mitochondrial dynamics links it to mitophagy pathways. These pathways regulate the removal of damaged mitochondria a vital process for cellular health and longevity.

MIRO1 has connections to neurodegenerative conditions such as Parkinson's disease and Alzheimer's disease. These conditions correlate with impaired mitochondrial trafficking and dynamics. MIRO1 interacts with proteins such as PINK1 and Parkin which contribute to mitochondrial quality control and are involved in the pathogenesis of these diseases. The disruption of MIRO1 function can lead to mitochondrial dysfunction and energy deficits which are underlying factors in these neurodegenerative processes.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Complementary Products

Primary Antibodies

AB320019

Anti-MIRO1 antibody [MJF-D29136-145] - BSA and Azide free (Detector)

Recombinant|RabMAb|KO Validated

Sandwich ELISA - Anti-MIRO1 antibody [MJF-D29136-145] - BSA and Azide free (Detector) (AB320019)

  • 0
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Primary Antibodies

AB321803

Anti-MIRO2 antibody [EPR29118-10]

BOND RX™ Validated|20ul selling size|Recombinant|RabMAb|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MIRO2 antibody [EPR29118-10] (AB321803)

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Cell Lines & Lysates

AB265792

Human RHOT1 (MIRO1) knockout HeLa cell line

Advanced Validation

Western blot - Human RHOT1 (MIRO1) knockout HeLa cell line (AB265792)

  • 0
  • 0
Proteins & Peptides

AB163047

Recombinant Human MIRO1 protein

SDS-PAGE - Recombinant Human MIRO1 protein (AB163047)

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Product promise

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