Human LAMB1 (Laminin beta 1) knockout HeLa cell lysate (ab257499)
Overview
-
Product name
Human LAMB1 (Laminin beta 1) knockout HeLa cell lysate -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
HeLa -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, Insertion of the selection cassette in exon3. -
Passage number
<20 -
Knockout validation
Sanger Sequencing, Western Blot (WB) -
Reconstitution notes
To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
-
Notes
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 - found here. 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.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.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.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.
-
Tested applications
Suitable for: WBmore details
Properties
-
Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab262952 - Human LAMB1 knockout HeLa cell lysate 1 x 100µg ab255929 - Human wild-type HeLa cell lysate 1 x 100µg -
Research areas
-
Cell type
epithelial -
Disease
Adenocarcinoma -
Gender
Female -
STR Analysis
Amelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8,12 CSF1PO: 9, 10
Target
-
Function
Binding to cells via a high affinity receptor, laminin is thought to mediate the attachment, migration and organization of cells into tissues during embryonic development by interacting with other extracellular matrix components. -
Sequence similarities
Contains 13 laminin EGF-like domains.
Contains 1 laminin IV type B domain.
Contains 1 laminin N-terminal domain. -
Domain
The alpha-helical domains I and II are thought to interact with other laminin chains to form a coiled coil structure.
Domains VI and IV are globular. -
Cellular localization
Secreted > extracellular space > extracellular matrix > basement membrane. Major component. - Information by UniProt
-
Alternative names
- CLM
- Cutis laxa with marfanoid phenotype
- LAM B1
see all
Associated products
-
KO cell lines
-
Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab257499 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB |
Use at an assay dependent concentration. Predicted molecular weight: 198 kDa.
|
| Notes |
|---|
|
WB
Use at an assay dependent concentration. Predicted molecular weight: 198 kDa. |
Images
-
Western blot - Human LAMB1 (Laminin beta 1) knockout HeLa cell lysate (ab257499)Lane 1: Wild-type HeLa cell lysate (20µg)
Lane 2: LAMB1 knockout HeLa cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - ab109293 observed at 200 kDa. Red - loading control ab7291 observed at 50 kDa.
ab109293 Anti-Laminin beta 1 antibody [EPR3189(2)] was shown to specifically 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 and Laminin beta 1 knockout samples 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) were incubated overnight at 4°C at 1 in 1000 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. -
Western blot - Human LAMB1 (Laminin beta 1) knockout HeLa cell lysate (ab257499)Lane 1: Wild-type HeLa cell lysate (20µg)
Lane 2: LAMB1 knockout HeLa cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - ab108536 observed at 200 kDa. Red - loading control ab7291 observed at 50 kDa.
ab108536 Anti-Laminin beta 1 antibody [EPR3189(2)] was shown to specifically 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 and Laminin beta 1 knockout samples 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) 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. -
Sanger Sequencing - Human LAMB1 knockout HeLa cell lysate (ab257499)Homozygous: Insertion of the selection cassette in exon3
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
-
SDS download
-
Datasheet download
References (0)
ab257499 has not yet been referenced specifically in any publications.