Human TMEM175 knockout HeLa cell lysate
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- WB
Lab
Western blot - Human TMEM175 knockout HeLa cell lysate (AB258722)
Anti-TMEM175 antibody [EPR24415-47] (ab300457) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, 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 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] (<a href='/en-us/products/primary-antibodies/tmem175-antibody-epr24415-47-ab300457'>ab300457</a>) 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
Secondary
Lanes 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
Predicted band size: 56 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human TMEM175 knockout HeLa cell lysate (AB258722)
Allele-1 : 1 bp insertion in exon4
Complementary Products
Primary Antibodies
AB300457
Anti-TMEM175 antibody [EPR24415-47]
20ul selling size|RabMAb|Recombinant|KO Validated
![Western blot - Anti-TMEM175 antibody [EPR24415-47] (AB300457)](https://content.abcam.com/products/images/tmem175-antibody-epr24415-47-ab300457--western-blot-img416707.jpg)
- 0
- 0
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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