Human PTBP1 knockout HeLa cell line
- Advanced Validation
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PTBP1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 4. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
57 kDa RNA-binding protein PPTB-1, HNRNP-I, HNRPI, Heterogeneous Nuclear Ribonucleoprotein Polypeptide I, Heterogeneous nuclear ribonucleoprotein I, MGC10830, MGC8461, PTB, PTB 1, PTB 2, PTB 3, PTB 4, PTB T, PTBP1_HUMAN, Polypyrimidine tract binding protein, Polypyrimidine tract binding protein (heterogeneous nuclear ribonucleoprotein I), Polypyrimidine tract-binding protein 1, RNA Binding Protein, pPTB
- WB
Lab
Western blot - Human PTBP1 knockout HeLa cell line (AB265155)
Lanes 1- 2 : Merged signal (red and green). Green - ab133734 observed at 57 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab133734 was shown to react with PTBP1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265155 (knockout cell lysate ab257614) was used. Wild-type HeLa and PTBP1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab133734 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 10000 dilution and a 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-PTBP1 antibody [EPR9048(B)] (<a href='/en-us/products/primary-antibodies/ptbp1-antibody-epr9048b-ab133734'>ab133734</a>) at 1/10000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
PTBP1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human PTBP1 knockout HeLa cell line (ab265155)
Predicted band size: 57 kDa
Observed band size: 57 kDa
false
- WB
Lab
Western blot - Human PTBP1 knockout HeLa cell line (AB265155)
Lanes 1- 2 : Merged signal (red and green). Green - ab134950 observed at 57 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab134950 was shown to react with PTBP1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265155 (knockout cell lysate ab257614) was used. Wild-type HeLa and PTBP1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab134950 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 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-PTBP1 antibody [EPR9049(B)] (<a href='/en-us/products/primary-antibodies/ptbp1-antibody-epr9049b-ab134950'>ab134950</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
PTBP1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human PTBP1 knockout HeLa cell line (ab265155)
Predicted band size: 57 kDa
Observed band size: 57 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human PTBP1 knockout HeLa cell line (AB265155)
Homozygous : 1 bp insertion in exon 4.
Reactivity data
Product details
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
PTBP1 influences the stability transport and translation of mRNA molecules. It is part of the heterogeneous nuclear ribonucleoprotein complex which plays a fundamental role in the processing and modification of pre-mRNA into mature mRNA. This activity impacts the expression of a wide range of genes affecting cellular function and growth. PTBP1 regulates mRNA metabolism contributing significantly to post-transcriptional gene regulation.
Pathways
The involvement of PTBP1 spans the management of alternative splicing and the modulation of gene expression. It participates importantly in the alternative splicing pathway alongside proteins like PTBP2 which perform overlapping functions in neurons. PTBP1 affects exon inclusion or exclusion influencing the genetic diversity presented in protein forms within cells. It also plays a role in the exon junction complex pathway impacting post-translational modifications and protein interactions.
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
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Cell Lines & Lysates
AB264958
Human CASP8 (Caspase-8) knockout HeLa cell line
Advanced Validation
- 0
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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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