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AB275248

Human GRB2 knockout HCT116 cell lysate

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GRB2 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in allele 1 and 2 bp deletion in allele 2 in exon 2.
5 Images
Western blot - Human GRB2 knockout HCT116 cell lysate (AB275248)
  • WB

Lab

Western blot - Human GRB2 knockout HCT116 cell lysate (AB275248)

Lane 1 : Wild type HCT116 (human colorectal carcinoma epithelial cell), whole cell lysate, 20 ug

Lane 2 : GRB2 knockout HCT116 (ab273715) whole cell lysate, 20 ug

Lane 3 : HeLa (human epithelial cell line from cervix adenocarcinoma), whole cell lysate, 20 ug

Lane 4 : 293T ( human embryonic kidney epithelial cell), whole cell lysate, 20 ug

Lanes 1-4 : Merged signal (red and green). Green - ab281846 observed at 26KDa. Red - loading control ab181602 (Rabbit monoclonal [EPR16891] to GAPDH) observed at 36 kDa.

Lanes 1-2 : ab281846 Anti-GRB2 antibody was shown to react with GRB2 in HCT116 cells in Western blot. Loss of signal was observed when GRB2 knockout cell line ab273715 (knockout cell lysate ab275248) was used. Wild-type and GRB2 knockout samples were subjected to SDS-PAGE.

ab281846 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated at 4℃ overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680CW) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800RD) preadsorbed (ab216772) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS

All lanes:

Western blot - Anti-GRB2 antibody [81/GRB2] (<a href='/en-us/products/primary-antibodies/grb2-antibody-81-grb2-ab281846'>ab281846</a>) at 1/1000 dilution

Lane 1:

Wild type HCT116 (human colorectal carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

GRB2 knockout HCT116 (<a href='/en-us/products/cell-lines/human-grb2-knockout-hct116-cell-line-ab273715'>ab273715</a>) whole cell lysate at 20 µg

Lane 3:

HeLa (human epithelial cell line from cervix adenocarcinoma), whole cell lysate at 20 µg

Lane 4:

293T ( human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Mouse IgG H&L (IRDye® 800CW) (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-800cw-preadsorbed-ab216772'>ab216772</a>) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution

Predicted band size: 25 kDa

Observed band size: 26 kDa

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Western blot - Human GRB2 knockout HCT116 cell lysate (AB275248)
  • WB

Unknown

Western blot - Human GRB2 knockout HCT116 cell lysate (AB275248)

Lane 1 : Wild-type HCT116 cell lysate 20 ug
Lane 2 : GRB2 knockout HCT116 cell lysate 20 ug
Lane 3 : HeLa cell lysate 20 ug
Lane 4 : HEK293 cell lysate 20 ug

Lanes 1 - 4 : Merged signal (red and green). Green - ab32111 observed at 25 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab32111 was shown to react with GRB2 in wild-type HCT 116 cells in western blot with loss of signal observed in GRB2 knockout cell line ab273715 (GRB2 knockout cell lysate ab275248). HCT 116 wild-type and GRB2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab32111 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 2000 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 hour at room temperature before imaging.

All lanes:

Western blot - Anti-GRB2 antibody [Y301] (<a href='/en-us/products/primary-antibodies/grb2-antibody-y301-ab32111'>ab32111</a>) at 1/2000 dilution

Lane 1:

Wild-type HCT116 cell lysate at 20 µg

Lane 2:

GRB2 knockout HCT116 cell lysate at 20 µg

Lane 2:

Western blot - Human GRB2 knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-grb2-knockout-hct116-cell-line-ab273715'>ab273715</a>)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

HEK293 cell lysate at 20 µg

Predicted band size: 25 kDa

Observed band size: 25 kDa

false

Sanger Sequencing - Human GRB2 knockout HCT116 cell lysate (AB275248)
  • Sanger seq

Unknown

Sanger Sequencing - Human GRB2 knockout HCT116 cell lysate (AB275248)

Allele-1 : 2 bp deletion in exon 2 .

Sanger Sequencing - Human GRB2 knockout HCT116 cell lysate (AB275248)
  • Sanger seq

Unknown

Sanger Sequencing - Human GRB2 knockout HCT116 cell lysate (AB275248)

Allele-2 : 1 bp deletion in exon 2 .

Western blot - Human GRB2 knockout HCT116 cell lysate (AB275248)
  • WB

Supplier Data

Western blot - Human GRB2 knockout HCT116 cell lysate (AB275248)

All lanes:

Western blot - Anti-GRB2 antibody [EPR24634-17] (<a href='/en-us/products/primary-antibodies/grb2-antibody-epr24634-17-ab315005'>ab315005</a>) at 1/1000 dilution

Lane 1:

Wild-type HCT116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Western blot - Human GRB2 knockout HCT116 cell lysate (ab275248) at 20 µg

Lane 3:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 4:

Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 6:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 7:

PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

Lane 8:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 25 kDa,36 kDa

false

Exposure time: 180s

Key facts

Cell type

HCT116

Species or organism

Human

Tissue

Colon

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in allele 1 and 2 bp deletion in allele 2 in exon 2

Disease

Carcinoma

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Complementary Products

Primary Antibodies

AB32037

Anti-GRB2 antibody [Y237]

RabMAb|Recombinant

Immunocytochemistry/ Immunofluorescence - Anti-GRB2 antibody [Y237] (AB32037)

  • 4
  • 3
Proteins & Peptides

AB280753

Recombinant Human PD1 protein

Mass Spectrometry - Recombinant Human PD1 protein (AB280753)

  • 0
  • 0
Proteins & Peptides

AB134868

Recombinant human CT-1 protein

Bioactive

  • 0
  • 0
Proteins & Peptides

AB217407

Recombinant human Vitronectin/S-Protein (Animal Free)

  • 0
  • 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 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
GRB2
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
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.

The GRB2 protein also known as growth factor receptor-bound protein 2 is instrumental in signal transduction. GRB2 has a molecular weight of about 25 kDa. It comprises one SH2 domain and two SH3 domains which facilitate its role in linking receptor tyrosine kinases to downstream signaling molecules. GRB2 is ubiquitously expressed in various tissues signifying its importance in diverse cellular functions.
Biological function summary

GRB2 functions as an adaptor protein that connects activated receptor tyrosine kinases to intracellular pathways. It often forms complexes with other proteins such as the SOS protein which further propagates signaling cascades critical for cell proliferation and differentiation. GRB2’s ability to mediate these interactions contributes to cellular responses to external stimuli including growth factors and hormones.

Pathways

GRB2 plays an important role in the Ras-MAPK signaling pathway. It interacts with proteins like SOS and Ras enabling signal transduction from the cell surface to the nucleus which is vital for processes such as cell growth and survival. GRB2 is also implicated in the PI3K-Akt pathway connecting it to another set of signaling proteins that regulate metabolism growth and survival.

GRB2 has significant associations with cancer and immune disorders. Aberrant activation of pathways involving GRB2 can lead to uncontrolled cell proliferation contributing to oncogenesis in different cancers. Additionally GRB2’s interactions with proteins like BCR-ABL in chronic myeloid leukemia highlight its potential role as a therapeutic target. GRB2 inhibitors could therefore offer promising avenues for treating such conditions by disrupting its pathological signaling interactions.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

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Product protocols

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Product promise

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For full details, please see our Terms & Conditions

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