Human TMED10 (TMP21) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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TMED10 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp insertion in exon 1.
View Alternative Names
1110014C03Rik, 21 kDa transmembrane-trafficking protein, MGC102351, P23, S31I125, S31III125, TMED 10, TMEDA_HUMAN, Tmed10, Tmp-21-I, Transmembrane emp24 domain-containing protein 10, Transmembrane emp24-like trafficking protein 10 (yeast), Transmembrane protein Tmp21, Transmembrane trafficking protein 21kD, p24 family protein delta-1, p24(DELTA), p24delta, p24delta1
- WB
Lab
Western blot - Human TMED10 (TMP21) knockout HEK-293T cell line (AB266227)
Lanes 1-4 : Merged signal (red and green). Green - ab134948 observed at 19 kDa. Red - loading control ab8245 observed at 36 kDa.
ab134948 Anti-TMP21 antibody [EPR9036(B)] was shown to specifically react with TMP21 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266227 (knockout cell lysate ab258233) was used. Wild-type and TMP21 knockout samples were subjected to SDS-PAGE. ab134948 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-TMP21 antibody [EPR9036(B)] (<a href='/en-us/products/primary-antibodies/tmp21-antibody-epr9036b-ab134948'>ab134948</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
TMED10 knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human TMED10 (TMP21) knockout HEK-293T cell line (ab266227)
Lane 3:
U-87 MG cell lysate at 20 µg
Lane 4:
Raji cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 25 kDa,9 kDa
Observed band size: 13 kDa,19 kDa
false
- WB
Lab
Western blot - Human TMED10 (TMP21) knockout HEK-293T cell line (AB266227)
Lanes 1-4 : Merged signal (red and green). Green - ab133771 observed at 19 kDa. Red - loading control ab8245 observed at 36 kDa.
ab133771 Anti-TMP21 antibody [EPR9037(B)] was shown to specifically react with TMP21 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266227 (knockout cell lysate ab258233) was used. Wild-type and TMP21 knockout samples were subjected to SDS-PAGE. ab133771 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-TMP21 antibody [EPR9037(B)] (<a href='/en-us/products/primary-antibodies/tmp21-antibody-epr9037b-ab133771'>ab133771</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
TMED10 knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human TMED10 (TMP21) knockout HEK-293T cell line (ab266227)
Lane 3:
U-87 MG cell lysate at 20 µg
Lane 4:
Raji cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 25 kDa,27 kDa,37 kDa,46 kDa
Observed band size: 19 kDa,27 kDa
false
- Cell Culture
Lab
Cell Culture - Human TMED10 (TMP21) knockout HEK-293T cell line (AB266227)
Representative images TMED10 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS M5000 microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human TMED10 (TMP21) knockout HEK-293T cell line (AB266227)
Homozygous : 2 bp insertion in exon1
Reactivity data
Product details
Recommended control: Human wild-type HEK293T cell line (ab255449). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
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
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
TMP21 affects the modulation of the gamma-secretase complex which is essential in the generation of amyloid precursor protein (APP) and its subsequent processing. This protein acts as a negative regulator of gamma-secretase activity leading to alterations in APP cleavage. By being part of this complex TMP21 exerts influence over processes such as signal transduction and protein metabolism. Its expression levels and activity can impact neurophysiological functions due to its presence in nervous tissues.
Pathways
TMP21 integrates into the amyloidogenic pathway influencing amyloid-beta peptide production. This pathway is associated with the APP processing where TMP21 regulates the cleavage by controlling gamma-secretase activity. Another pathway involving TMP21 is the protein transport pathway facilitating the orderly movement of proteins through the early secretory pathway. Within these frameworks TMP21 interacts with proteins like presenilin which is a significant component of the gamma-secretase complex thereby affecting its overall functionality.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB134948
Anti-TMP21 antibody [EPR9036(B)]
RabMAb|Recombinant|KO Validated|Trial Size
![Western blot - Anti-TMP21 antibody [EPR9036(B)] (AB134948)](https://content.abcam.com/products/images/tmp21-antibody-epr9036b-ab134948--western-blot-img272.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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