AB265414

Human LAMB1 (Laminin beta 1) knockout HeLa cell line

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LAMB1 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: Insertion of the selection cassette in exon 3. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

別名を表示する

CLM, Cutis laxa with marfanoid phenotype, LAMB1_HUMAN, LIS5, Laminin B1, Laminin B1 chain, Laminin beta 1 chain, Laminin beta 1 chain precursor, Laminin beta1, Laminin subunit beta-1, Laminin-1 subunit beta, Laminin-10 subunit beta, Laminin-12 subunit beta, Laminin-2 subunit beta, Laminin-6 subunit beta, Laminin-8 subunit beta, MGC142015

3 Images

Lab

Western blot - Human LAMB1 (Laminin beta 1) knockout HeLa cell line (AB265414)

Lanes 1- 2 : Merged signal (red and green). Green - ab108536 observed at 200 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

ab108536 was shown to react with Laminin beta 1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265414 (knockout cell lysate ab257499) was used. Wild-type HeLa and LAMB1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab108536 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4° at a 1 in 1000 dilution and a 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-Laminin beta 1 antibody [EPR3189(2)] (<a href='/products/primary-antibodies/laminin-beta-1-antibody-epr31892-ab108536'>ab108536</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 1:

Western blot - Human LAMB1 (Laminin beta 1) knockout HeLa cell lysate (<a href='/products/cell-lysates/human-lamb1-laminin-beta-1-knockout-hela-cell-lysate-ab257499'>ab257499</a>)

Lane 2:

LAMB1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human LAMB1 (Laminin beta 1) knockout HeLa cell line (ab265414)

Predicted band size: 198 kDa

Observed band size: 200 kDa

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Lab

Western blot - Human LAMB1 (Laminin beta 1) knockout HeLa cell line (AB265414)

Lanes 1- 2 : Merged signal (red and green). Green - ab109293 observed at 200 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

ab109293 was shown to react with Laminin beta 1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265414 (knockout cell lysate ab257499) was used. Wild-type HeLa and LAMB1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109293 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4° at a 1 in 1000 dilution and a 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-Laminin beta 1 antibody [EPR3190(2)] (<a href='/products/primary-antibodies/laminin-beta-1-antibody-epr31902-ab109293'>ab109293</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

LAMB1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human LAMB1 (Laminin beta 1) knockout HeLa cell line (ab265414)

Predicted band size: 198 kDa

Observed band size: 200 kDa

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Sanger Sequencing - Human LAMB1 (Laminin beta 1) knockout HeLa cell line (AB265414)

Sanger seq

Unknown

Sanger Sequencing - Human LAMB1 (Laminin beta 1) knockout HeLa cell line (AB265414)

Homozygous : Insertion of the selection cassette in exon 3.

Key facts

細胞タイプ

HeLa

生物種

Human

組織

Cervix

製品の状態

Liquid

form

ノックアウト検証方法

Sanger Sequencing,Western blot

ノックアウト変異

Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 3

疾病

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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製品の詳細

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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製品内容

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出荷温度及び保存条件

遺伝子名

LAMB1

遺伝子編集のタイプ

Knockout

遺伝子編集の方法

CRISPR technology

ノックアウト検証方法

Sanger Sequencing, Western blot

接合型

Homozygous

出荷温度

Dry Ice

短期保存温度

-196°C

長期保存温度

-196°C

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取り扱い方法

初回取り扱いガイドライン

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.

継代培養ガイドライン

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.

培養培地

DMEM (High Glucose) + 10% FBS

凍結保存培地

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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補足情報

This supplementary information is collated from multiple sources and compiled automatically.

Laminin beta 1 also known as LAMB1 is a subunit of the laminin molecule an essential component of the extracellular matrix. This protein has an approximate molecular weight of 200 kDa. Laminin beta 1 is one of the three chains that form the laminin heterotrimer. It is predominantly expressed in basement membranes which provide structural support and segregate tissues. The laminin 1 (laminin-111) complex including the LAMB1 chain contributes to cell adhesion differentiation migration and plays a role in tissue development and repair.

Biological function summary

This component of the laminin molecule plays a critical role in maintaining the integrity of tissues by organizing cell architecture and mediating cellular signals. Laminin beta 1 is part of a larger laminin protein complex that interacts with a variety of cell surface receptors including integrins and dystroglycan contributing to cell communication and signal transduction. These interactions guide cell orientation and regulate vital processes such as cell movement and growth.

Pathways

Laminin beta 1 participates in the integrin signaling pathway and the PI3K-AKT signaling pathway. It interacts closely with integrin proteins such as integrin alpha-6 and integrin beta-4 during these processes. In integrin signaling the laminin molecule facilitates the binding between cells and their surrounding extracellular matrix. This connection influences cellular responses including survival and motility which are necessary for normal development and wound healing processes.

Alterations in laminin beta 1 expression are linked to conditions like muscular dystrophy and Pierson syndrome. In muscular dystrophy the dysfunction of laminin beta 1 interacts with proteins such as dystrophin in muscle cells contributing to compromised structural integrity and muscle wasting. Similarly in Pierson syndrome abnormalities in laminin beta 1 are associated with a rare genetic disorder that affects the kidneys and eyes where it implicates the protein nephrin in the glomerular basement membrane leading to serious renal dysfunction.

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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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製品プロトコール

Visit the General protocols
Visit the Troubleshooting

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Abcam product promise

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Human LAMB1 (Laminin beta 1) knockout HeLa cell line