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AB265677

Human ANXA6 (Annexin-6) knockout HeLa cell line

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ANXA6 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 7 and 5 bp deletion in exon 7.

View Alternative Names

67 kDa calelectrin, ANX 6, ANXA6_HUMAN, Annexin A6, Annexin VI, Annexin VI p68, Annexin-6, CBP 68, CPB-II, Calcium binding protein p68, Calelectrin, Calphobindin-II, Chromobindin-20, Lipocortin VI, P68, Protein III, p70

5 Images
Western blot - Human ANXA6 (Annexin-6) knockout HeLa cell line (AB265677)
  • WB

Lab

Western blot - Human ANXA6 (Annexin-6) knockout HeLa cell line (AB265677)

Lanes 1-4 : Merged signal (red and green). Green - ab201023 observed at 75 kDa. Red - loading control ab8245 observed at 36 kDa.

ab201023 Anti-Annexin-6/ANXA6 antibody [EPR19536] was shown to specifically react with 6/ANXA6 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265677 (knockout cell lysate ab257351) was used. Wild-type and 6/ANXA6 knockout samples were subjected to SDS-PAGE. ab201023 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours 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-Annexin-6/ANXA6 antibody [EPR19517] (<a href='/en-us/products/primary-antibodies/annexin-6-anxa6-antibody-epr19517-ab201023'>ab201023</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

ANXA6 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human ANXA6 (Annexin-6) knockout HeLa cell line (ab265677)

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

Raji 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: 76 kDa

Observed band size: 75 kDa

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Western blot - Human ANXA6 (Annexin-6) knockout HeLa cell line (AB265677)
  • WB

Lab

Western blot - Human ANXA6 (Annexin-6) knockout HeLa cell line (AB265677)

Lanes 1 - 4 : Merged signal (red and green). Green - ab199422 observed at 75 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab199422 was shown to react with Annexin-6/ANXA6 in wild-type HeLa cells in western blot with loss of signal observed in ANXA6 knockout cell line ab265677 (ANXA6 knockout cell lysate ab257351). Wild-type and ANXA6 knockout HeLa cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab199422 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4° at a 1 in 1000 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-Annexin-6/ANXA6 antibody [EPR17308] - N-terminal (<a href='/en-us/products/primary-antibodies/annexin-6-anxa6-antibody-epr17308-n-terminal-ab199422'>ab199422</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

ANXA6 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human ANXA6 (Annexin-6) knockout HeLa cell line (ab265677)

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

Raji cell lysate at 20 µg

Predicted band size: 76 kDa

Observed band size: 75 kDa

false

Western blot - Human ANXA6 (Annexin-6) knockout HeLa cell line (AB265677)
  • WB

Lab

Western blot - Human ANXA6 (Annexin-6) knockout HeLa cell line (AB265677)

Lanes 1-4 : Merged signal (red and green). Green - ab201024 observed at 75 kDa. Red - loading control ab8245 observed at 36 kDa.

ab201024 Anti-Annexin-6/ANXA6 antibody [EPR19536] was shown to specifically react with Annexin-6/ANXA6 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265677 (knockout cell lysate ab257351) was used. Wild-type and Annexin-6/ANXA6 knockout samples were subjected to SDS-PAGE. ab201024 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours 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-Annexin-6/ANXA6 antibody [EPR19536] (<a href='/en-us/products/primary-antibodies/annexin-6-anxa6-antibody-epr19536-ab201024'>ab201024</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

ANXA6 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human ANXA6 (Annexin-6) knockout HeLa cell line (ab265677)

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

Raji 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: 76 kDa

Observed band size: 75 kDa

false

Sanger Sequencing - Human ANXA6 (Annexin-6) knockout HeLa cell line (AB265677)
  • Sanger seq

Unknown

Sanger Sequencing - Human ANXA6 (Annexin-6) knockout HeLa cell line (AB265677)

Allele-2 : 1 bp insertion in exon 7.

Sanger Sequencing - Human ANXA6 (Annexin-6) knockout HeLa cell line (AB265677)
  • Sanger seq

Unknown

Sanger Sequencing - Human ANXA6 (Annexin-6) knockout HeLa cell line (AB265677)

Allele-1 : 5 bp deletion in exon 7.

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, 1 bp insertion in exon 7 and 5 bp deletion in exon 7

Antibiotic resistance

Puromycin 1µg/mL

Disease

Adenocarcinoma

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Reactivity data

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Product details

Recommended control: Human wild-type HeLa cell line (ab255928). 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.

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What's included?

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Properties and storage information

Gene name
ANXA6
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
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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.

Annexin-6 also known as ANXA6 is a member of the annexin family of proteins. It has a mass of approximately 68 kDa and is expressed in various tissues with substantial presence in muscle and liver. This protein acts mechanically by binding to membranes in a calcium-dependent manner playing a role in membrane-associated processes such as endocytosis and exocytosis. Annexin-6 can also interact with cytoskeletal elements influencing cell structure and stability.
Biological function summary

Annexin-6 functions as a versatile scaffold that organizes multiprotein complexes. It modulates membrane trafficking processes and participates in signaling pathways within the cell. This protein can influence intracellular calcium homeostasis by interacting with calcium channels and transporters making it an integral part of cellular signaling and communication.

Pathways

Annexin-6 plays a role in the MAPK and PI3K/Akt signaling pathways. It associates with proteins such as Ras and Protein Kinase C impacting cell proliferation survival and differentiation. These interactions highlight the importance of annexin-6 in cellular responses to external stimuli and help regulate various physiological processes in the body.

Annexin-6 has been linked to cardiac disorders and cancer. In the context of cardiac health it interacts with proteins like phospholamban and calmodulin to influence heart muscle function and calcium regulation. In cancer annexin-6's involvement in the Akt signaling pathway can affect tumor growth and metastasis often interacting with other proteins like EGFR and HER2. This makes it a significant protein of interest in disease research and potential therapeutic targeting.
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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 ANXA6
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