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AB266646

Human COPS7B (CSN7b) knockout HEK-293T cell line

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Human COPS7B (CSN7b) knockout HEK-293T cell line available to order. Recommended control: Human wild-type HEK293T cell line (ab255449).

View Alternative Names

COP9 Complex Homolog Subunit 7b, COP9 Constitutive Photomorphogenic Homolog Subunit 7b, COP9 Signalasome Subunit 7b, COP9 signalosome complex subunit 7b, COPS7B, CSN7B_HUMAN, JAB1-containing signalosome subunit 7b, SGN7b, Signalosome subunit 7b

3 Images
Western blot - Human COPS7B (CSN7b) knockout HEK-293T cell line (AB266646)
  • WB

Lab

Western blot - Human COPS7B (CSN7b) knockout HEK-293T cell line (AB266646)

Lanes 1-4 : Merged signal (red and green). Green - ab124718 observed at 32 kDa. Red - loading control ab8245 observed at 36 kDa.

ab124718 Anti-CSN7b antibody [EPR6465] was shown to specifically react with CSN7b in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266646 (knockout cell lysate ab257895) was used. Wild-type and CSN7b knockout samples were subjected to SDS-PAGE. ab124718 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° 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-CSN7b antibody [EPR6465] (<a href='/en-us/products/primary-antibodies/csn7b-antibody-epr6465-ab124718'>ab124718</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

COPS7B knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human COPS7B (CSN7b) knockout HEK-293T cell line (ab266646)

Lane 3:

HeLa 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: 30 kDa

Observed band size: 32 kDa

false

Western blot - Human COPS7B (CSN7b) knockout HEK-293T cell line (AB266646)
  • WB

Lab

Western blot - Human COPS7B (CSN7b) knockout HEK-293T cell line (AB266646)

Lanes 1-4 : Merged signal (red and green). Green - ab133548 observed at 32 kDa. Red - loading control ab8245 observed at 36 kDa.

ab133548 Anti-CSN7b antibody [EPR6464] was shown to specifically react with CSN7b in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266646 (knockout cell lysate ab257895) was used. Wild-type and CSN7b knockout samples were subjected to SDS-PAGE. ab133548 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° 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-CSN7b antibody [EPR6464] (<a href='/en-us/products/primary-antibodies/csn7b-antibody-epr6464-ab133548'>ab133548</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

COPS7B knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human COPS7B (CSN7b) knockout HEK-293T cell line (ab266646)

Lane 3:

HeLa 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: 30 kDa

Observed band size: 32 kDa

false

Sanger Sequencing - Human COPS7B (CSN7b) knockout HEK-293T cell line (AB266646)
  • Sanger seq

Unknown

Sanger Sequencing - Human COPS7B (CSN7b) knockout HEK-293T cell line (AB266646)

Homozygous : 1 bp insertion in exon3

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: 1 bp insertion in exon 3

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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, 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.

What's included?

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Properties and storage information

Gene name
COPS7B
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

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.

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

For this product, it's our understanding that no specific protocols are required. You can visit:

Product promise

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