TMEM175 KO cell lysate available now. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon4 and Insertion of the selection cassette in exon4.
Images
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 exon4 and Insertion of the selection cassette in exon4.
Alternative names
Hypothetical protein DKFZp547K246, Hypothetical protein MGC4618, MGC4618, TM175_HUMAN, Transmembrane protein 175, hypothetical protein LOC84286
What's included?
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TMEM175 KO cell lysate available now. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon4 and Insertion of the selection cassette in exon4.
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 exon4 and Insertion of the selection cassette in exon4.
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- TMEM175
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Female
Storage
- Shipped at conditions
- Ambient - Can Ship with Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
Notes
Knockout cell lysate achieved by CRISPR/Cas9.
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
TMEM175 also known as Transmembrane Protein 175 is a channel protein with dual function as a lysosomal potassium channel. It has an approximate mass of 66 kDa. TMEM175 plays a role in ion homeostasis and is expressed mainly in tissues with high levels of lysosomes including the brain and the immune system. This protein aids in the maintenance of the lysosomal environment important for cellular activities.
- Biological function summary
This lysosomal protein contributes to the maintenance of ion stability within lysosomes an important factor in lysosomal degradation and overall cellular clearance processes. TMEM175 is an integral component of the lysosomal membrane and participates in ensuring lysosomal pH stability therefore influencing autophagic activity. TMEM175 does not participate in known protein complexes but it is pivotal for preserving lysosomal integrity.
- Pathways
TMEM175 actively engages in the lysosomal degradation pathway impacting cellular waste processing. TMEM175 influences endolysosomal trafficking and autophagy which are essential for metabolite recycling. It works alongside proteins like ATPase and lysosomal enzyme hydrolases by controlling ion concentrations which can influence other lysosomal functions and pathways.
- Associated diseases and disorders
TMEM175 has a significant connection with neurodegenerative conditions such as Parkinson's disease. Mutation or dysfunction in TMEM175 impacts lysosomal function linked to protein aggregation often involving alpha-synuclein which serves as a hallmark for such neurodegenerative diseases. Additionally TMEM175 is implicated in lysosomal storage disorders where impaired lysosomal function results in cellular accumulation of substances due to ineffective degradation.
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2 product images
Sanger Sequencing - Human TMEM175 knockout HeLa cell lysate (ab258722)
Allele-1: 1 bp insertion in exon4
Western blot - Human TMEM175 knockout HeLa cell lysate (ab258722)
Anti-TMEM175 antibody [EPR24415-47] (Anti-TMEM175 antibody [EPR24415-47] ab300457) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-TMEM175 antibody [EPR24415-47] ab300457 was shown to bind specifically to TMEM175 only when the lysate samples were not boiled. A band was observed at 40-50 kDa in wild-type HeLa cell lysates with no signal observed at this size in TMEM175 knockout cell line Human TMEM175 knockout HeLa cell line ab265571 (knockout cell lysate ab258722). To generate this image, wild-type and TMEM175 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 5 % 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-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
Lane 1: Wild-type HeLa (boiled) cell lysate, 20 ug
Lane 2: TMEM175 knockout HeLa (boiled) cell lysate, 20 ug
Lane 3: U-87 MG (boiled) cell lysate, 20 ug
Lane 4: Wild-type HeLa (unboiled) cell lysate, 20 ug
Lane 5: TMEM175 knockout HeLa (unboiled) cell lysate, 20 ug
Lane 6: U-87 MG (unboiled) cell lysate, 20 ug
Observed MV band: 40-50kDa.All lanes: Western blot - Anti-TMEM175 antibody [EPR24415-47] (Anti-TMEM175 antibody [EPR24415-47] ab300457) at 1/1000 dilution
Lane 1: Wild-type HeLa (boiled) cell lysate at 20 µg
Lane 2: TMEM175 knockout HeLa (boiled) cell lysate at 20 µg
Lane 3: U-87 MG (boiled) cell lysate at 20 µg
Lane 4: Wild-type HeLa (unboiled) cell lysate at 20 µg
Lane 5: TMEM175 knockout HeLa (unboiled) cell lysate at 20 µg
Lane 6: U-87 MG (unboiled) cell lysate at 20 µg
SecondaryLanes 1 - 6: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 6: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 56 kDa
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