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AB264819

Human ITGA2 (CD49b) knockout HeLa cell line

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ITGA2 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: 14 bp deletion in exon 4. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
4 Images
Western blot - Human ITGA2 (CD49b) knockout HeLa cell line (AB264819)
  • WB

Lab

Western blot - Human ITGA2 (CD49b) knockout HeLa cell line (AB264819)

Lanes 1-4 : Merged signal (red and green). Green - ab181549 observed at 160 kDa. Red - loading control ab8245 observed at 37 kDa.

ab181549 Anti-Integrin alpha 2 antibody [EPR17349] was shown to specifically react with Integrin alpha 2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264819 (knockout cell lysate ab257262) was used. Wild-type and Integrin alpha 2 knockout samples were subjected to SDS-PAGE. ab181549 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-Integrin alpha 2 antibody [EPR17349] (<a href='/en-us/products/primary-antibodies/integrin-alpha-2-antibody-epr17349-ab181549'>ab181549</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

ITGA2 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human ITGA2 (CD49b) knockout HeLa cell line (ab264819)

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

K562 cell lysate at 20 µg

Predicted band size: 129 kDa,130 kDa,171 kDa

Observed band size: 160 kDa,250 kDa

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Western blot - Human ITGA2 (CD49b) knockout HeLa cell line (AB264819)
  • WB

Lab

Western blot - Human ITGA2 (CD49b) knockout HeLa cell line (AB264819)

Lanes 1-4 : Merged signal (red and green). Green - ab109432 observed at 129 kDa. Red - loading control ab8245 observed at 37 kDa.

ab109432 Anti-Integrin alpha 2 antibody [EPR5789] was shown to specifically react with Integrin alpha 2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264819 (knockout cell lysate ab257262) was used. Wild-type and Integrin alpha 2 knockout samples were subjected to SDS-PAGE. ab109432 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-Integrin alpha 2 antibody [EPR5789] (<a href='/en-us/products/primary-antibodies/integrin-alpha-2-antibody-epr5789-ab109432'>ab109432</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

ITGA2 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human ITGA2 (CD49b) knockout HeLa cell line (ab264819)

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

K562 cell lysate at 20 µg

Predicted band size: 129 kDa,130 kDa

Observed band size: 129 kDa

false

Sanger Sequencing - Human ITGA2 (CD49b) knockout HeLa cell line (AB264819)
  • Sanger seq

Unknown

Sanger Sequencing - Human ITGA2 (CD49b) knockout HeLa cell line (AB264819)

Homozygous : 14 bp deletion in exon 4.

Cell Culture - Human ITGA2 (CD49b) knockout HeLa cell line (AB264819)
  • Cell Culture

Unknown

Cell Culture - Human ITGA2 (CD49b) knockout HeLa cell line (AB264819)

Representative images of ITGA2 knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.

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: 14 bp deletion in exon 4

Antibiotic resistance

Puromycin 1µg/mL

Disease

Adenocarcinoma

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Reactivity data

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

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What's included?

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Properties and storage information

Gene name
ITGA2
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.

CD49b also known as Integrin alpha-2 or ITGA2 is a transmembrane glycoprotein with a molecular weight of approximately 160 kDa. This protein localizes on the surface of various cells including platelets fibroblasts and some immune cells. It functions as part of the integrin family which are receptors that mediate the cell-extracellular matrix (ECM) interactions. CD49b binds to collagen and laminin in the ECM and contributes to cellular adhesion and signal transduction.
Biological function summary

The CD49b protein plays a critical role in cell migration thrombosis and wound healing. By forming a heterodimer with Integrin beta-1 it becomes a part of the collagen receptor complex Integrin alpha-2/beta-1 which facilitates various cellular interactions with the ECM. This functionality enables cells to sense their environment and respond to mechanical changes which is important for processes such as tissue repair and embryonic development.

Pathways

Several cellular mechanisms involve CD49b in maintaining physiological homeostasis. One significant pathway is the integrin signaling pathway where CD49b interacts with focal adhesion kinase (FAK) and other intracellular proteins to regulate cytoskeletal rearrangement and cell motility. Another pathway is platelet activation where CD49b collaborates with glycoprotein VI (GPVI) and other receptors to initiate platelet aggregation during hemostasis. These pathways highlight the importance of CD49b in various cellular functions related to cell mobility and stability.

Researchers have linked CD49b to conditions including thrombosis and fibrosis. In thrombosis abnormal platelet aggregation often involves CD49b and its interaction with fibrinogen and von Willebrand factor. In fibrosis CD49b's role in cell adhesion and matrix remodeling correlates with excessive ECM deposition and scar formation. Understanding CD49b's function offers insights into therapeutic strategies alongside related proteins such as Integrin beta-1 which may modulate disease progression.
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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 ITGA2
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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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