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AB257351

Human ANXA6 (Annexin-6) knockout HeLa cell lysate

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ANXA6 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon7 and 5 bp deletion in exon7.

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 lysate (AB257351)
  • WB

Lab

Western blot - Human ANXA6 (Annexin-6) knockout HeLa cell lysate (AB257351)

Lane 1 : Wild-type HeLa cell lysate (20 ug)
Lane 2 : ANXA6 knockout HeLa cell lysate (20 ug)
Lane 3 : Jurkat cell lysate (20 ug)
Lane 4 : Raji cell lysate (20 ug)

ab201023 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. 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 (<a href='/en-us/products/cell-lines/human-anxa6-annexin-6-knockout-hela-cell-line-ab265677'>ab265677</a>)

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

Western blot - Human ANXA6 (Annexin-6) knockout HeLa cell lysate (AB257351)
  • WB

Lab

Western blot - Human ANXA6 (Annexin-6) knockout HeLa cell lysate (AB257351)

Lane 1 : Wild-type HeLa cell lysate 20 ug
Lane 2 : ANXA6 knockout HeLa cell lysate 20 ug
Lane 3 : Jurkat cell lysate 20 ug
Lane 4 : Raji cell lysate 20 ug
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°C 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 (<a href='/en-us/products/cell-lines/human-anxa6-annexin-6-knockout-hela-cell-line-ab265677'>ab265677</a>)

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 lysate (AB257351)
  • WB

Lab

Western blot - Human ANXA6 (Annexin-6) knockout HeLa cell lysate (AB257351)

Lane 1 : Wild-type HeLa cell lysate (20 ug)
Lane 2 : ANXA6 knockout HeLa cell lysate (20 ug)
Lane 3 : Jurkat cell lysate (20 ug)
Lane 4 : Raji cell lysate (20 ug)

ab201024 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 (<a href='/en-us/products/cell-lines/human-anxa6-annexin-6-knockout-hela-cell-line-ab265677'>ab265677</a>)

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 lysate (AB257351)
  • Sanger seq

Unknown

Sanger Sequencing - Human ANXA6 (Annexin-6) knockout HeLa cell lysate (AB257351)

Allele-2 : 1 bp insertion in exon7

Sanger Sequencing - Human ANXA6 (Annexin-6) knockout HeLa cell lysate (AB257351)
  • Sanger seq

Unknown

Sanger Sequencing - Human ANXA6 (Annexin-6) knockout HeLa cell lysate (AB257351)

Allele-1 : 5 bp deletion in exon7

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon7 and 5 bp deletion in exon7.

Disease

Adenocarcinoma

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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab257351-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human ANXA6 knockout HeLa cell lysate", "number":"AB257351-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HeLa cell lysate", "number":"AB257351-CMP02", "productcode":"" } ] } } }
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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
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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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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Product promise

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