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AB266530

Human PLCG1 (Phospholipase C gamma 1/PLC-gamma-1) knockout HEK-293T cell line

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PLCG1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 267 bp deletion in exon 1.

View Alternative Names

1 phosphatidyl D myo inositol 4 5 bisphosphate, 1-phosphatidylinositol-4,5-bisphosphate phosphodiesterase gamma-1, Inositoltrisphosphohydrolase, Monophosphatidylinositol phosphodiesterase, NCKAP3, PLC 1, PLC-148, PLC-II, PLC-gamma-1, PLCG1_HUMAN, Phosphatidylinositol phospholipase C, Phosphoinositidase C, Phosphoinositide phospholipase C, Phosphoinositide phospholipase C-gamma-1, Phospholipase C 148, Phospholipase C-II, Phospholipase C-gamma-1

3 Images
Western blot - Human PLCG1 (Phospholipase C gamma 1/PLC-gamma-1) knockout HEK-293T cell line (AB266530)
  • WB

Lab

Western blot - Human PLCG1 (Phospholipase C gamma 1/PLC-gamma-1) knockout HEK-293T cell line (AB266530)

Lanes 1- 2 : Merged signal (red and green). Green - ab109501 observed at 160 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab109501 was shown to react with Phospholipase C gamma 1/PLC-gamma-1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266530 (knockout cell lysate ab257590) was used. Wild-type HEK-293T and PLCG1 knockout HEK-293T 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. ab109501 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° 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-Phospholipase C gamma 1/PLC-gamma-1 antibody [EPR5358] (<a href='/en-us/products/primary-antibodies/phospholipase-c-gamma-1-plc-gamma-1-antibody-epr5358-ab109501'>ab109501</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 40 µg/mL

Lane 2:

PLCG1 knockout HEK-293T cell lysate at 40 µg/mL

Lane 2:

Western blot - Human PLCG1 (Phospholipase C gamma 1/PLC-gamma-1) knockout HEK-293T cell line (ab266530)

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 1/20000 dilution

Predicted band size: 148 kDa

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Sanger Sequencing - Human PLCG1 (Phospholipase C gamma 1/PLC-gamma-1) knockout HEK-293T cell line (AB266530)
  • Sanger seq

Unknown

Sanger Sequencing - Human PLCG1 (Phospholipase C gamma 1/PLC-gamma-1) knockout HEK-293T cell line (AB266530)

Homozygous : 267 bp deletion in exon1

Immunocytochemistry/ Immunofluorescence - Human PLCG1 (Phospholipase C gamma 1/PLC-gamma-1) knockout HEK-293T cell line (AB266530)
  • ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Human PLCG1 (Phospholipase C gamma 1/PLC-gamma-1) knockout HEK-293T cell line (AB266530)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized PLCG1 KO HEK293T (ab266530) cells labelling Phospholipase Cγ1 with ab302940 at 1/100 (10.63 ug/ml) dilution, followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/mL dilution (Green). Confocal image showing cytoplasmic staining in parental HEK293T cell line and no staining in PLCG1 KO HEK293T cell line is observed. ab202272 Recombinant Alexa Fluor® 594 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab202272) was used to counterstain tubulin at 1/50 10 ug/mL dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 267 bp deletion in exon 1

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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

Recommended control: Human wild-type HEK293T cell line (ab255449). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

We will provide viable cells that proliferate on revival.

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
PLCG1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
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.

Phospholipase C gamma 1 (PLC-gamma-1) also known as PLCG1 or PLC-gamma plays an important role in cellular signaling. This enzyme has a molecular mass of approximately 150 kDa. It is found in multiple cell types but is highly expressed in the brain and immune cells. PLC-gamma-1 is involved in the hydrolysis of phosphatidylinositol 45-bisphosphate (PIP2) to generate inositol 145-trisphosphate (IP3) and diacylglycerol (DAG) leading to downstream signaling events that control various cellular processes.
Biological function summary

PLC-gamma-1 is integral in signal transduction and is an important player in the immune response. The protein does not function as part of a large multi-protein complex but interacts transiently with other signaling molecules. It acts downstream of growth factor receptors and immunoreceptors by mediating calcium release and activation of protein kinase C influencing cell proliferation migration and differentiation. Its expression pattern suggests a critical role in the nervous and immune systems affecting learning memory and immune defense.

Pathways

PLC-gamma-1 is an important component in both the phospholipase C and PLC-gamma pathways. These pathways include important interactions with proteins such as the receptor tyrosine kinases and Src family kinases. In these pathways PLC-gamma-1 modulates signals that guide cellular responses to external stimuli affecting processes like growth signaling and T-cell receptor activation which is vital in adaptive immunity.

PLC-gamma-1 has significant connections to cancer and immune deficiencies. Abnormal PLC-gamma-1 activity is observed in certain cancers such as breast cancer where it influences tumor growth and metastasis. Its dysregulation in immune cells leads to disorders like autoimmune diseases by impacting proteins such as LAT and SLP-76 which are important in T-cell signaling. Understanding PLC-gamma-1's regulatory mechanisms helps in developing targeted therapies for these conditions.
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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 PLCG1
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