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AB265026

Human CALD1 (Caldesmon/CDM) knockout HeLa cell line

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CALD1 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 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
3 Images
Western blot - Human CALD1 (Caldesmon/CDM) knockout HeLa cell line (AB265026)
  • WB

Lab

Western blot - Human CALD1 (Caldesmon/CDM) knockout HeLa cell line (AB265026)

Lanes 1-4 : Merged signal (red and green). Green - ab183339 observed at 75 kDa. Red - loading control ab8245 observed at 36 kDa.

ab183339 Anti-Caldesmon/CDM antibody [SP226] was shown to specifically react with Caldesmon/CDM in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265026 (knockout cell lysate ab257375) was used. Wild-type and Caldesmon/CDM knockout samples were subjected to SDS-PAGE. ab183339 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-Caldesmon/CDM antibody [SP226] (<a href='/en-us/products/primary-antibodies/caldesmon-cdm-antibody-sp226-ab183339'>ab183339</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CALD1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CALD1 (Caldesmon/CDM) knockout HeLa cell line (ab265026)

Lane 3:

NIH/3T3 cell lysate at 20 µg

Lane 4:

HEK-293 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: 93 kDa

Observed band size: 75 kDa

false

Western blot - Human CALD1 (Caldesmon/CDM) knockout HeLa cell line (AB265026)
  • WB

Lab

Western blot - Human CALD1 (Caldesmon/CDM) knockout HeLa cell line (AB265026)

Lanes 1-4 : Merged signal (red and green). Green - ab32330 observed at 75 kDa. Red - loading control ab8245 observed at 36 kDa.

ab32330 Anti-Caldesmon/CDM antibody [E89] was shown to specifically react with Caldesmon/CDM in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265026 (knockout cell lysate ab257375) was used. Wild-type and Caldesmon/CDM knockout samples were subjected to SDS-PAGE. ab32330 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-Caldesmon/CDM antibody [E89] (<a href='/en-us/products/primary-antibodies/caldesmon-cdm-antibody-e89-ab32330'>ab32330</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CALD1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CALD1 (Caldesmon/CDM) knockout HeLa cell line (ab265026)

Lane 3:

NIH/3T3 cell lysate at 20 µg

Lane 4:

HEK-293 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: 93 kDa

Observed band size: 75 kDa

false

Sanger Sequencing - Human CALD1 (Caldesmon/CDM) knockout HeLa cell line (AB265026)
  • Sanger seq

Unknown

Sanger Sequencing - Human CALD1 (Caldesmon/CDM) knockout HeLa cell line (AB265026)

Homozygous : 1 bp insertion in exon 1.

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 1

Antibiotic resistance

Puromycin 1µg/mL

Disease

Adenocarcinoma

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Complementary Products

Primary Antibodies

AB32330

Anti-Caldesmon/CDM antibody [E89]

BOND RX™ Validated|RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caldesmon/CDM antibody [E89] (AB32330)

  • 0
  • 0
Proteins & Peptides

AB313361

Recombinant Human WFDC2 Protein (His tag)

Mass Spectrometry - Recombinant Human WFDC2 Protein (His tag) (AB313361)

  • 0
  • 0
Proteins & Peptides

AB217407

Recombinant human Vitronectin/S-Protein (Animal Free)

  • 0
  • 0
Proteins & Peptides

AB317877

Recombinant Human UCH-L1/PGP9.5 Protein (His-tag)

Mass Spectrometry - Recombinant Human UCH-L1/PGP9.5 Protein (His-tag) (AB317877)

  • 0
  • 0
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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
CALD1
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.

Caldesmon also known as CDM CALD1 and h-caldesmon is a cytoskeletal protein with a molecular mass of approximately 93 to 120 kDa. This protein expresses itself abundantly in smooth muscle tissues yet it can also be found in non-muscle cells like fibroblasts and endothelial cells. Researchers often use specific caldesmon antibodies in immunohistochemistry allowing them to label various cellular components and providing insights into tissue composition. Due to its presence in muscle tissues caldesmon is essential for understanding muscle physiology and pathology.
Biological function summary

Caldesmon interacts with actin and myosin to regulate actomyosin contractility. This protein plays a critical role in controlling the contraction and relaxation processes in smooth muscle cells. Caldesmon forms part of a complex that includes calmodulin and tropomyosin enhancing its ability to stabilize actin filaments. It functions by inhibiting the ATPase activity of myosin therefore influencing cellular motility and shape change mechanisms. Researchers continually study caldesmon to comprehend its interactome and its significance within the larger cellular structure.

Pathways

Caldesmon participates in the regulation of the cytoskeletal dynamics vital for cell motility and structural integrity. In particular it is an important component of the contraction-relaxation cycle pathway in smooth muscle tissues. This protein has connections with pathways involving RhoA-Rho kinase where caldesmon modulates the phosphorylation levels influencing muscle contraction. Additionally proteins like tropomyosin and calmodulin modulate its activity especially under calcium-calmodulin-dependent pathways which further elucidates its regulatory importance.

Caldesmon has associations with conditions like certain types of cancers and cardiovascular diseases. Its expression levels and distribution provide valuable information in identifying smooth muscle tumors and other pathological conditions. In oncology for example h-caldesmon serves as a marker to distinguish leiomyosarcomas from other tumors. Moreover due to its involvement in smooth muscle contractility caldesmon links with proteins such as calmodulin and tropomyosin in diseases where abnormal contraction and cellular motility play significant roles. Understanding these connections is important for developing targeted treatments and improving diagnostic accuracy.
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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 CALD1
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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

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