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AB258586

Human PKN1 knockout HEK-293T cell lysate

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PKN1 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 insertion in exon2 and Insertion of the selection cassette in exon2.
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Western blot - Human PKN1 knockout HEK-293T cell lysate (AB258586)
  • WB

Lab

Western blot - Human PKN1 knockout HEK-293T cell lysate (AB258586)

Lane 1 : Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 2 : PKN1 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 3 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate (20 ug)
Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate (20 ug)

ab108976 was shown to specifically react with PKN in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266599 (knockout cell lysate ab258586) was used. Wild-type and PKN knockout samples were subjected to SDS-PAGE. ab108976 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4oC 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-PKN antibody [EPR3237] (<a href='/en-us/products/primary-antibodies/pkn-antibody-epr3237-ab108976'>ab108976</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

PKN1 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

Western blot - Human PKN1 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-pkn1-knockout-hek-293t-cell-line-ab266599'>ab266599</a>)

Lane 3:

SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate at 20 µg

Lane 4:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole 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: 104 kDa

Observed band size: 120 kDa

false

Sanger Sequencing - Human PKN1 knockout HEK-293T cell lysate (AB258586)
  • Sanger seq

Unknown

Sanger Sequencing - Human PKN1 knockout HEK-293T cell lysate (AB258586)

Allele-2 : Insertion of the selection cassette in exon2

Sanger Sequencing - Human PKN1 knockout HEK-293T cell lysate (AB258586)
  • Sanger seq

Unknown

Sanger Sequencing - Human PKN1 knockout HEK-293T cell lysate (AB258586)

Allele-1 : 1 bp insertion in exon2

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon2 and Insertion of the selection cassette in exon2.

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Complementary Products

Primary Antibodies

AB124709

Anti-PKN3 (phospho T718) + PRK1 (phospho T774) + PKN2 (phospho T816) antibody [EPR5671]

RabMAb|Recombinant|Trial Size

Western blot - Anti-PKN3 (phospho T718) + PRK1 (phospho T774) + PKN2 (phospho T816) antibody [EPR5671] (AB124709)

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Proteins & Peptides

AB269032

Recombinant human GPSN2-PKN1 fusion protein (Tagged)

Functional Studies - Recombinant human GPSN2-PKN1 fusion protein (Tagged) (AB269032)

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Cell Lines & Lysates

AB266599

Human PKN1 knockout HEK-293T cell line

Advanced Validation

Western blot - Human PKN1 knockout HEK-293T cell line (AB266599)

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  • 0
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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></p>" } } }
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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.

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What's included?

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Properties and storage information

Gene name
PKN1
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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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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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com