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AB257235

Human GRN (Granulin) knockout HEK-293T cell lysate

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GRN KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon2 and 2 bp deletion in exon2.
4 Images
Western blot - Human GRN (Granulin) knockout HEK-293T cell lysate (AB257235)
  • WB

Lab

Western blot - Human GRN (Granulin) knockout HEK-293T cell lysate (AB257235)

All lanes:

Western blot - Anti-Progranulin antibody [EPR15864] (<a href='/en-us/products/primary-antibodies/granulin-antibody-epr15864-ab208777'>ab208777</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T lysate at 20 µg

Lane 2:

Western blot - Human GRN (Granulin) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-grn-granulin-knockout-hek-293t-cell-line-ab266738'>ab266738</a>)

Lane 2:

Western blot - Human GRN (Granulin) knockout HEK-293T cell lysate (ab257235)

Observed band size: 64 kDa

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Western blot - Human GRN (Granulin) knockout HEK-293T cell lysate (AB257235)
  • WB

Lab

Western blot - Human GRN (Granulin) knockout HEK-293T cell lysate (AB257235)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : GRN knockout HEK293T cell lysate (20 ug)
Lane 3 : HepG2 cell lysate (20 ug)
Lane 4 : HAP1cell lysate (20 ug)

ab208777 was shown to specifically react with Progranulin in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266738 (knockout cell lysate ab257235) was used. Wild-type and Progranulin knockout samples were subjected to SDS-PAGE. ab208777 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-Progranulin antibody [EPR15864] (<a href='/en-us/products/primary-antibodies/granulin-antibody-epr15864-ab208777'>ab208777</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

GRN knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human GRN (Granulin) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-grn-granulin-knockout-hek-293t-cell-line-ab266738'>ab266738</a>)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

HAP1 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: 64 kDa

Observed band size: 74 kDa

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Sanger Sequencing - Human GRN (Granulin) knockout HEK-293T cell lysate (AB257235)
  • Sanger seq

Unknown

Sanger Sequencing - Human GRN (Granulin) knockout HEK-293T cell lysate (AB257235)

Allele-1 : 14 bp deletion in exon2

Sanger Sequencing - Human GRN (Granulin) knockout HEK-293T cell lysate (AB257235)
  • Sanger seq

Unknown

Sanger Sequencing - Human GRN (Granulin) knockout HEK-293T cell lysate (AB257235)

Allele-2 : 2 bp deletion 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, 14 bp deletion in exon2 and 2 bp deletion in exon2.

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Complementary Products

Cell Lines & Lysates

AB266738

Human GRN (Granulin) knockout HEK-293T cell line

Advanced Validation

Western blot - Human GRN (Granulin) knockout HEK-293T cell line (AB266738)

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Primary Antibodies

AB208777

Anti-Progranulin antibody [EPR15864]

20ul selling size|RabMAb|Recombinant|KO Validated|Trial Size

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Progranulin antibody [EPR15864] (AB208777)

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Assay Kits

AB252364

Human Progranulin ELISA Kit

Recombinant|SimpleStep

Sandwich ELISA - Human Progranulin ELISA Kit (AB252364)

  • 0
  • 0
Assay Kits

AB219524

Mouse Progranulin Matched Antibody Pair Kit

Recombinant

ELISA - Mouse Progranulin Matched Antibody Pair Kit (AB219524)

  • 5
  • 1
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Reactivity data

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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 and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant 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
GRN
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.

Granulin also known as progranulin or GRN is a protein with a molecular mass of approximately 68 kDa. This protein is encoded by the GRN gene and is involved in various cellular processes. Granulin is expressed widely within the body particularly in epithelial cells immune cells and neurons. Researchers have developed methods like GRN ELISA and progranulin ELISA to quantitatively measure the levels of granulin in biological samples aiding in further understanding of its role in health and disease.
Biological function summary

Granulin functions as a growth factor involved in tissue repair cell proliferation and inflammation. It does not function as part of a larger protein complex but acts individually in exerting its effects on cells. Granulin influences various aspects of cellular architecture and signaling pathways making it important for maintaining cellular homeostasis. Additionally the granulin protein participates in regulating networks that involve cytokines and growth factors impacting both the innate and adaptive immune responses.

Pathways

Granulin plays significant roles in the MAPK and NF-kB signaling pathways. These pathways critically manage cellular reactions to stress inflammation and other external stimuli. Granulin modulates these processes often interacting with proteins like TNF receptors and interleukins to affect the downstream signaling events. Interactions between granulin and other signaling molecules allow it to integrate into broader networks that oversee various biological responses.

Significant links exist between granulin mutations and frontotemporal dementia (FTD) and Alzheimer's disease. Mutations in the GRN gene often lead to reduced granulin levels contributing to neurodegenerative conditions. These mutations affect the protein's normal function promoting pathological mechanisms associated with neuronal deterioration. Granulin interacts with proteins such as TDP-43 in neurodegenerative diseases where alterations might result in protein aggregation and neurotoxicity. Understanding these connections can lead to potential therapeutic interventions targeting granulin-related pathways.
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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'.

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