Human TGFB1 (TGF beta 1) knockout A549 cell lysate (ab269671)
Overview
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Product name
Human TGFB1 (TGF beta 1) knockout A549 cell lysate
See all TGF beta 1 kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9.
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Parental Cell Line
A549 -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift = 99.45% -
Passage number
<20 -
Knockout validation
Next Generation Sequencing (NGS), 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.
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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.
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Tested applications
Suitable for: WBmore details
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab280546 - Human TGFB1 knockout A549cell lysate 1 x 100µg ab259782 - Human wild-type A549 cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
Disease
Carcinoma -
Gender
Male
Target
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Function
Multifunctional protein that controls proliferation, differentiation and other functions in many cell types. Many cells synthesize TGFB1 and have specific receptors for it. It positively and negatively regulates many other growth factors. It plays an important role in bone remodeling as it is a potent stimulator of osteoblastic bone formation, causing chemotaxis, proliferation and differentiation in committed osteoblasts. -
Tissue specificity
Highly expressed in bone. Abundantly expressed in articular cartilage and chondrocytes and is increased in osteoarthritis (OA). Co-localizes with ASPN in chondrocytes within OA lesions of articular cartilage. -
Involvement in disease
Defects in TGFB1 are the cause of Camurati-Engelmann disease (CE) [MIM:131300]; also known as progressive diaphyseal dysplasia 1 (DPD1). CE is an autosomal dominant disorder characterized by hyperostosis and sclerosis of the diaphyses of long bones. The disease typically presents in early childhood with pain, muscular weakness and waddling gait, and in some cases other features such as exophthalmos, facial paralysis, hearing difficulties and loss of vision. -
Sequence similarities
Belongs to the TGF-beta family. -
Post-translational
modificationsGlycosylated.
The precursor is cleaved into mature TGF-beta-1 and LAP, which remains non-covalently linked to mature TGF-beta-1 rendering it inactive. -
Cellular localization
Secreted > extracellular space > extracellular matrix. - Information by UniProt
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Alternative names
- Cartilage-inducing factor
- CED
- Differentiation inhibiting factor
see all
Associated products
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KO cell lines
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab269671 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.
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| Notes |
|---|
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WB
Use at an assay dependent concentration. |
Images
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Western blot - Human TGFB1 (TGF beta 1) knockout A549 cell lysate (ab269671)
Lane 1: Wild-type A549 cell lysate 20 μg
Lane 2: TGFB1 knockout A549 cell lysate 20 μg
Lane 3: K562 cell lysate 20 μg
Lane 4: SH-SY5Y cell lysate 20 μgLanes 1 - 4: Merged signal (red and green). Green - ab215715 observed at 48 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab215715 was shown to react with TGF beta 1 in wild-type A549 cells in Western blot with loss of signal observed in TGFB1 knockout cell line ab269509 (TGFB1 knockout cell lysate ab269671). Wild-type A549 and TGFB1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab215715 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) 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 h at room temperature before imaging.
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Western blot - Human TGFB1 (TGF beta 1) knockout A549 cell lysate (ab269671)Lane 1: Wild-type A549 cell lysate 20 μg
Lane 2: TGFB1 knockout A549 cell lysate 20 μg
Lane 3: K562 cell lysate 20 μg
Lane 4: SH-SY5Y cell lysate 20 μgLanes 1 - 4: Merged signal (red and green). Green - ab179695 observed at 48 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab179695 was shown to react with TGF beta in wild-type A549 cells in Western blot with loss of signal observed in TGFB1 knockout cell line ab269509 (TGFB1 knockout cell lysate ab269671). Wild-type A549 and TGFB1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab179695 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) 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 h at room temperature before imaging.
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
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SDS download
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Datasheet download
References (0)
ab269671 has not yet been referenced specifically in any publications.