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AB266033

Human Plcb1 (Phospholipase C beta 1) knockout HeLa cell line

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PLCB1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 5. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

View Alternative Names

1 phosphatidylinositol 4,5 bisphosphate phosphodiesterase beta 1, 1-phosphatidyl-D-myo-inositol-4,5-bisphosphate, 1-phosphatidylinositol-4, 5-bisphosphate phosphodiesterase beta-1, EIEE12, Inositoltrisphosphohydrolase, Monophosphatidylinositol phosphodiesterase, PI PLC, PLC 1, PLC-154, PLC-I, PLC-beta-1, PLCB1_HUMAN, Phosphb, Phosphoinositidase C, Phosphoinositide phospholipase C, Phosphoinositide phospholipase C-beta-1, Phospholipase C beta 1 (phosphoinositide-specific), Phospholipase C-I, Phospholipase C-beta-1, Plcb1 protein, Triphosphoinositide phosphodiesterase

2 Images
Western blot - Human Plcb1 (Phospholipase C beta 1) knockout HeLa cell line (AB266033)
  • WB

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Western blot - Human Plcb1 (Phospholipase C beta 1) knockout HeLa cell line (AB266033)

Lanes 1-4 : Merged signal (red and green). Green - ab182359 observed at 150 kDa. Red - loading control ab8245 observed at 36 kDa.

ab182359 Anti-Phospholipase C beta 1/PLCB1 antibody [EPR19085] was shown to specifically react with Phospholipase C beta 1/PLCB1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab266033 (knockout cell lysate ab257589) was used. Wild-type and Phospholipase C beta 1/PLCB1 knockout samples were subjected to SDS-PAGE. ab182359 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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-Phospholipase C beta 1/PLCB1 antibody [EPR19085] (<a href='/en-us/products/primary-antibodies/phospholipase-c-beta-1-plcb1-antibody-epr19085-ab182359'>ab182359</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PLCB1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human Plcb1 (Phospholipase C beta 1) knockout HeLa cell line (ab266033)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

Mouse heart tissue 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: 139 kDa

Observed band size: 150 kDa

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Sanger Sequencing - Human Plcb1 (Phospholipase C beta 1) knockout HeLa cell line (AB266033)
  • Sanger seq

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Sanger Sequencing - Human Plcb1 (Phospholipase C beta 1) knockout HeLa cell line (AB266033)

Homozygous : 1 bp insertion in exon5

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 5

Disease

Adenocarcinoma

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Reactivity data

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Product details

We will provide viable cells that proliferate on revival.

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
PLCB1
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 beta 1 (PLCB1) also known as PLC-β1 or PLCB1 is an enzyme involved in intracellular signal transduction. With a molecular weight of around 150 kDa PLCB1 hydrolyzes phosphatidylinositol 45-bisphosphate (PIP2) into inositol trisphosphate (IP3) and diacylglycerol (DAG). This biochemical activity plays an important role in the release of calcium ions from intracellular stores. PLCB1 is expressed in regions of the brain including the cerebral cortex as well as in other tissues.
Biological function summary

Multiple functions depend on the signaling cascades initiated by PLCB1. It belongs to a larger family of phospholipase C enzymes contributing to the regulation of various neural processes. The enzyme acts within multiprotein complexes that modulate intracellular calcium levels influencing neuronal activity and synaptic transmission. This role implicates PLCB1 in modulating brain functions related to learning and memory.

Pathways

PLCB1 contributes to cellular signaling pathways such as the phosphoinositide signaling pathway and the G protein-coupled receptor (GPCR) pathway. In these pathways PLCB1 interacts with major proteins such as Gαq subunit of G proteins which activates it. These interactions place PLCB1 as a central player in the pathways that help convert extracellular signals into intracellular actions often involving the regulation of secondary messengers like IP3 and DAG.

Researchers associate PLCB1 with neurological and psychiatric conditions including epilepsy and bipolar disorder. Dysregulation in PLCB1 activity links to altered calcium signaling which can affect brain function. Additionally proteins like Gαq and other GPCR pathway elements may either buffer or exacerbate effects arising from PLCB1 anomalies suggesting molecular interdependencies in related disease mechanisms.
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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 PLCB1
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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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