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AB257617

Human PTPN1 (PTP1B) knockout HeLa cell lysate

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PTPN1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. 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 lysate (AB257617)
  • WB

Lab

Western blot - Human PTPN1 (PTP1B) knockout HeLa cell lysate (AB257617)

Lane 1 : Wild-type HeLa cell lysate 20 μg
Lane 2 : PTPN1 knockout Hela cell lysate 20 μg
Lane 3 : Jurkat cell lysate 20 μg
Lane 4 : MCF7 cell lysate 20 μg
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 37kDa.
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 (<a href='/en-us/products/cell-lines/human-ptpn1-ptp1b-knockout-hela-cell-line-ab265014'>ab265014</a>)

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 lysate (AB257617)
  • WB

Unknown

Western blot - Human PTPN1 (PTP1B) knockout HeLa cell lysate (AB257617)

Lane 1 : Wild-type HeLa cell lysate (20µg)

Lane 2 : PTPN1 knockout HeLa cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab252928 observed at 50 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab252928 Anti-PTP1B antibody [EPR22468-6] was shown to specifically react with PTP1B in wild-type HeLa cells in western blot. The band observed in the 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 and PTP1B knockout samples 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 1 in 1000 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-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 knockout HeLa cell lysate at 20 µg

Predicted band size: 50 kDa

Observed band size: 50 kDa

false

Western blot - Human PTPN1 (PTP1B) knockout HeLa cell lysate (AB257617)
  • WB

Lab

Western blot - Human PTPN1 (PTP1B) knockout HeLa cell lysate (AB257617)

Lane 1 : Wild-type HeLa cell lysate (20µg)

Lane 2 : PTPN1 knockout HeLa cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab244207 observed at 50 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab244207 Anti-PTP1B antibody [EPR22474] was shown to specifically react with PTP1B in wild-type HeLa cells in western blot. The band observed in the 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 and PTP1B knockout samples 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 1 in 1000 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-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

Predicted band size: 50 kDa

Observed band size: 50 kDa

false

Western blot - Human PTPN1 (PTP1B) knockout HeLa cell lysate (AB257617)
  • WB

Lab

Western blot - Human PTPN1 (PTP1B) knockout HeLa cell lysate (AB257617)

Lane 1 : Wild-type HeLa cell lysate 20 μg
Lane 2 : PTPN1 knockout HeLa cell lysate 20 μg
Lane 3 : Jurkat cell lysate 20 μg
Lane 4 : MCF7 cell lysate 20 μg
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 37kDa.
ab244207 was shown to specifically react with PTP1B in wild-type HeLa cells in western blot. The band observed in the 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 (<a href='/en-us/products/cell-lines/human-ptpn1-ptp1b-knockout-hela-cell-line-ab265014'>ab265014</a>)

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 lysate (AB257617)
  • Sanger seq

Unknown

Sanger Sequencing - Human PTPN1 (PTP1B) knockout HeLa cell lysate (AB257617)

Allele-2 : 1 bp insertion in exon 4

Sanger Sequencing - Human PTPN1 (PTP1B) knockout HeLa cell lysate (AB257617)
  • Sanger seq

Unknown

Sanger Sequencing - Human PTPN1 (PTP1B) knockout HeLa cell lysate (AB257617)

Allele-1 : 1 bp deletion in exon 4

Sanger Sequencing - Human PTPN1 (PTP1B) knockout HeLa cell lysate (AB257617)
  • Sanger seq

Unknown

Sanger Sequencing - Human PTPN1 (PTP1B) knockout HeLa cell lysate (AB257617)

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

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

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

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

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.

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What's included?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab257617-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human PTPN1 knockout HeLa cell lysate", "number":"AB257617-CMP02", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HeLa cell lysate", "number":"AB257617-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
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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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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Complementary Products

Primary Antibodies

AB244207

Anti-PTP1B antibody [EPR22474]

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Proteins & Peptides

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Assay Kits

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Biochemicals

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