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AB266708

Human CECR5 knockout HEK-293T cell line

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HDHD5 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 17 bp deletion in exon 6 and 1 bp deletion in exon 6. 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

A930002G03Rik, CECR5_HUMAN, Cat eye syndrome chromosome region, candidate 5, Cat eye syndrome chromosome region, candidate 5 homolog, Cat eye syndrome critical region protein 5, MGC25951, OTTMUSP00000025905

4 Images
Western blot - Human CECR5 knockout HEK-293T cell line (AB266708)
  • WB

Lab

Western blot - Human CECR5 knockout HEK-293T cell line (AB266708)

Lanes 1-4 : Merged signal (red and green). Green - ab184185 observed at 46 kDa. Red - loading control ab8245 observed at 36 kDa.

ab184185 Anti-CECR5 antibody [EPR14793(B)] - C-terminal was shown to specifically react with CECR5 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266708 (knockout cell lysate ab257888) was used. Wild-type and CECR5 knockout samples were subjected to SDS-PAGE. ab184185 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-CECR5 antibody [EPR14793(B)] - C-terminal (<a href='/en-us/products/primary-antibodies/cecr5-antibody-epr14793b-c-terminal-ab184185'>ab184185</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

CECR5 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human CECR5 knockout HEK-293T cell line (ab266708)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

Jurkat 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: 46 kDa

Observed band size: 46 kDa

false

Cell Culture - Human CECR5 knockout HEK-293T cell line (AB266708)
  • Cell Culture

Unknown

Cell Culture - Human CECR5 knockout HEK-293T cell line (AB266708)

Representative images of CECR5 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using an EVOS M5000 microscope.

Sanger Sequencing - Human CECR5 knockout HEK-293T cell line (AB266708)
  • Sanger seq

Unknown

Sanger Sequencing - Human CECR5 knockout HEK-293T cell line (AB266708)

Allele-2 : 1 bp deletion in exon 6.

Sanger Sequencing - Human CECR5 knockout HEK-293T cell line (AB266708)
  • Sanger seq

Unknown

Sanger Sequencing - Human CECR5 knockout HEK-293T cell line (AB266708)

Allele-1 : 17 bp deletion in exon 6

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, 17 bp deletion in exon 6 and 1 bp deletion in exon 6

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 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.

What's included?

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Properties and storage information

Gene name
HDHD5
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

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.

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

CECR5 also known as Cat Eye Syndrome Critical Region Candidate 5 encodes a protein with a mass of about 48 kDa. This protein shows expression in multiple tissues such as the brain kidney and liver. Its mechanical role involves binding interactions that help in signaling and cellular communication processes. The protein's structure enables it to participate effectively in these interactions making it a point of interest for further study in these specific tissues.
Biological function summary

CECR5 serves an important function in cellular signaling networks and is not typically found as part of a larger complex. The protein plays a role in modulating certain cellular responses possibly impacting processes like differentiation and growth. Researchers have identified CECR5 involvement in cellular mechanisms that govern normal development and maintenance hinting at its broader influence in cellular homeostasis.

Pathways

CECR5 engages in the MAPK signaling pathway which is instrumental in transferring extracellular signals to intracellular responses. The MAPK pathway involves proteins like ERK and JNK with which CECR5 may interact or have regulatory effects. Additionally it influences signaling via the PI3K-AKT pathway a pathway that controls many aspects of cell survival growth and metabolism. This association suggests CECR5's potential roles in important cellular activities and regulatory mechanisms.

Alterations in CECR5 expression or function relate to Cat Eye Syndrome and possibly neurodevelopmental disorders. In Cat Eye Syndrome the overexpression or structural changes of CECR5 may disrupt normal development alongside chromosomal duplications involving the region. Connections also appear between CECR5 and other proteins like PAX6 in the context of its role in developmental processes. Understanding these relationships further explains CECR5's impact on these conditions.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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