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AB265014

Human PTPN1 (PTP1B) knockout HeLa cell line

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PTPN1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 4 and 1 bp insertion in exon 4 and Insertion of the selection cassette in exon 4.

View Alternative Names

Non receptor tyrosine phosphatase 1, PTN1_HUMAN, PTP-1B, PTPN 1, Protein phosphotyrosylphosphatase 1B, Protein tyrosine phosphatase non receptor type 1, Protein tyrosine phosphatase placental, Protein-tyrosine phosphatase 1B, Tyrosine-protein phosphatase non-receptor type 1

7 Images
Western blot - Human PTPN1 (PTP1B) knockout HeLa cell line (AB265014)
  • WB

Lab

Western blot - Human PTPN1 (PTP1B) knockout HeLa cell line (AB265014)

Lanes 1- 2 : Merged signal (red and green). Green - ab244207 observed at 50 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab244207 was shown to react with PTP1B in wild-type HeLa cells in western blot. The band observed in knockout cell line ab265014 (knockout cell lysate ab257617) lane below 50kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and PTPN1 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. ab244207 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated 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-PTP1B antibody [EPR22474] (<a href='/en-us/products/primary-antibodies/ptp1b-antibody-epr22474-ab244207'>ab244207</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PTPN1 CRISPR/Cas9 edited HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human PTPN1 (PTP1B) knockout HeLa cell line (ab265014)

Predicted band size: 50 kDa

Observed band size: 50 kDa

false

Western blot - Human PTPN1 (PTP1B) knockout HeLa cell line (AB265014)
  • WB

Lab

Western blot - Human PTPN1 (PTP1B) knockout HeLa cell line (AB265014)

Lanes 1- 2 : Merged signal (red and green). Green - ab252928 observed at 50 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab252928 was shown to react with PTP1B in wild-type HeLa cells in western blot. The band observed in knockout cell line ab265014 (knockout cell lysate ab257617) lane below 50kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and PTPN1 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. ab252928 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated 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-PTP1B antibody [EPR22468-6] (<a href='/en-us/products/primary-antibodies/ptp1b-antibody-epr22468-6-ab252928'>ab252928</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PTPN1 CRISPR/Cas9 edited HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human PTPN1 (PTP1B) knockout HeLa cell line (ab265014)

Predicted band size: 50 kDa

Observed band size: 50 kDa

false

Western blot - Human PTPN1 (PTP1B) knockout HeLa cell line (AB265014)
  • WB

Lab

Western blot - Human PTPN1 (PTP1B) knockout HeLa cell line (AB265014)

Lanes 1 - 4 : Merged signal (red and green). Green - ab244207 observed at 51 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab244207 was shown to react with PTP1B in wild-type HeLa cells in western blot. The band observed in knockout cell line ab265014 (knockout cell lysate ab257617) lane below 50kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and PTPN1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5 % milk in TBS-T (0.1 % Tween®) before incubation with ab244207 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-PTP1B antibody [EPR22474] (<a href='/en-us/products/primary-antibodies/ptp1b-antibody-epr22474-ab244207'>ab244207</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PTPN1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human PTPN1 (PTP1B) knockout HeLa cell line (ab265014)

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

MCF7 cell lysate at 20 µg

Predicted band size: 50 kDa

Observed band size: 51 kDa

false

Western blot - Human PTPN1 (PTP1B) knockout HeLa cell line (AB265014)
  • WB

Lab

Western blot - Human PTPN1 (PTP1B) knockout HeLa cell line (AB265014)

Lanes 1 - 4 : Merged signal (red and green). Green - ab201974 observed at 51 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37 kDa.

ab201974 was shown to react with PTP1B in wild-type HeLa cells in Western blot with loss of signal observed in PTPN1 knockout cell line ab265014 (PTPN1 knockout cell lysate ab257617). Wild-type HeLa and PTPN1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5 % milk in TBS-T (0.1 % Tween®) before incubation with ab201974 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4 °C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-PTP1B antibody [4F8F11] (<a href='/en-us/products/primary-antibodies/ptp1b-antibody-4f8f11-ab201974'>ab201974</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PTPN1 knockout Hela cell lysate at 20 µg

Lane 2:

Western blot - Human PTPN1 (PTP1B) knockout HeLa cell line (ab265014)

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

MCF7 cell lysate at 20 µg

Predicted band size: 50 kDa

Observed band size: 51 kDa

false

Sanger Sequencing - Human PTPN1 (PTP1B) knockout HeLa cell line (AB265014)
  • Sanger seq

Unknown

Sanger Sequencing - Human PTPN1 (PTP1B) knockout HeLa cell line (AB265014)

Allele-2 : 1 bp insertion in exon 4.

Sanger Sequencing - Human PTPN1 (PTP1B) knockout HeLa cell line (AB265014)
  • Sanger seq

Unknown

Sanger Sequencing - Human PTPN1 (PTP1B) knockout HeLa cell line (AB265014)

Allele-1 : 1 bp deletion in exon 4.

Sanger Sequencing - Human PTPN1 (PTP1B) knockout HeLa cell line (AB265014)
  • Sanger seq

Unknown

Sanger Sequencing - Human PTPN1 (PTP1B) knockout HeLa cell line (AB265014)

Allele-3 : Insertion of the selection cassette in exon 4.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 4 and 1 bp insertion in exon 4 and Insertion of the selection cassette in exon 4

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>Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.</p>" } } }
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Product details

We will provide viable cells that proliferate on revival.

Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.

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?

{ "values": { "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab265014-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab265014 Human PTPN1 (PTP1B) knockout HeLa cell line", "number":"AB265014-CMP01", "productcode":"" } ] } } }
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Properties and storage information

Gene name
PTPN1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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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.

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Supplementary information

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

Protein tyrosine phosphatase 1B (PTP1B) is an enzyme involved in the dephosphorylation of tyrosine residues in proteins. Known also as PTPN1 this protein has a molecular weight of approximately 50 kDa. PTP1B is widely expressed in tissues but is highly present in the liver muscle and adipose tissue. It plays a critical role in regulating signal transduction pathways by removing phosphate groups from phosphotyrosine residues on target proteins. This action generally serves as a 'turn-off' switch for various signaling pathways.
Biological function summary

PTP1B plays important roles in glucose and lipid metabolism. It is not generally considered a part of a complex functioning instead as a singular enzyme. Its main action involves the negative regulation of the insulin signaling pathway by dephosphorylating the insulin receptor as well as its associated substrates. This regulation helps maintain normal levels of insulin sensitivity influencing cellular glucose uptake and metabolism.

Pathways

PTP1B has important functions in the insulin and leptin signaling pathways. It has a regulatory influence in these pathways affecting key metabolic processes. In the insulin signaling pathway PTP1B closely interacts with the insulin receptor as well as with insulin receptor substrates (IRS). Its dephosphorylating activity can lead to decreased insulin sensitivity. In the leptin pathway PTP1B modulates leptin signaling by dephosphorylating the Janus kinase 2 (JAK2) another protein involved in the control of energy balance and metabolism.

Excessive activity of PTP1B links with disorders such as type 2 diabetes and obesity. Overactivity of PTP1B can lead to insulin resistance contributing to the pathogenesis of type 2 diabetes. In obesity PTP1B may interfere with the normal activity of leptin a hormone critical for regulating energy balance. Its interaction with the insulin receptor and JAK2 makes PTP1B a target for therapeutic interventions aimed at improving insulin sensitivity and energy homeostasis.
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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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Target data

See full target information PTPN1
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