TGM2 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 1 bp insertion 8 bp deletion Frameshift = 97%.
Images
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Mutation description
- Knockout achieved by CRISPR/Cas9 X = 1 bp insertion 8 bp deletion Frameshift = 97%
Alternative names
ALPHA SUBUNIT, C polypeptide, EC 2.3.2.13, G alpha h, GNAH, GNAH G PROTEIN, G[a]h, Gh CLASS G ALPHA h, H POLYPEPTIDE, HEL-S-45, Protein-glutamine gamma-glutamyltransferase 2, TG 2, TG(C), TGC, TGC GUANINE NUCLEOTIDE BINDING PROTEIN, TGM2_HUMAN, TGase C, TGase H, TGase-2, TgaseII, Tissue transglutaminase, Transglutaminase 2 C polypeptide, Transglutaminase C, Transglutaminase H, Transglutaminase-2, epididymis secretory protein Li 45, tTG, tTGas
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TGM2 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 1 bp insertion 8 bp deletion Frameshift = 97%.
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Mutation description
- Knockout achieved by CRISPR/Cas9 X = 1 bp insertion 8 bp deletion Frameshift = 97%
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- TGM2
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Male
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.
- Do not allow the cell density to exceed 7x104 cells/cm2.
- Culture medium
- F-12K + 10% FBS
- Cryopreservation medium
- Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Transglutaminase 2 also known as TG2 or tissue transglutaminase is an enzyme with a mass of approximately 74 kDa. It catalyzes the formation of covalent bonds by crosslinking glutamine residues and lysine residues in proteins. This enzyme is widely expressed in various tissues including the liver heart and lungs. It can act as a GTPase a kinase and a protein disulfide isomerase highlighting its multifunctional nature. Additionally transglutaminase 2 associates with cell membranes cytoskeleton and extracellular matrix indicating its versatile involvement in cellular processes.
- Biological function summary
Transglutaminase 2 plays roles in cell adhesion wound healing and the immune response. It mediates extracellular matrix stabilization through crosslinking structural proteins which is critical for tissue repair. The enzyme does not typically form a stable part of a large complex but interacts with fibronectin as a binding partner highlighting its role in cellular adhesion and migration. The enzymatic activity of transglutaminase 2 helps maintain tissue integrity by modifying extracellular and intracellular proteins.
- Pathways
Transglutaminase 2 contributes significantly to the integrin-mediated signaling pathway and the apoptosis pathway. It assists in the modulation of cell-matrix interactions by crosslinking matrix proteins which is important for integrin signaling. During apoptosis transglutaminase 2 promotes cellular processes that lead to cell death via its involvement in nuclear condensation and DNA fragmentation. In these pathways proteins such as fibronectin and integrins interact with transglutaminase 2 facilitating its regulatory functions.
- Associated diseases and disorders
Transglutaminase 2 is associated with celiac disease and neurodegenerative disorders like Huntington's disease. Its autoantigenic properties make it a target in celiac disease where antibodies against transglutaminase 2 play a role in the pathology by affecting intestinal tissue. In Huntington's disease transglutaminase 2 contributes to the formation of protein aggregates which are toxic to neurons. This enzyme's interactions with proteins such as gliadin in celiac disease and huntingtin in Huntington's disease are central to its pathogenic mechanism.
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9 product images
Western blot - Human TGM2 (Transglutaminase 2) knockout A549 cell line (ab261876)
Lanes 1 - 4: Merged signal (red and green). Green - Anti-Transglutaminase 2 antibody [CUB 7402] ab2386 observed at 77 kDa. Red - loading control Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 observed at 37 kDa.Anti-Transglutaminase 2 antibody [CUB 7402] ab2386 was shown to recognize in wild-type A549 cells as signal was lost at the expected MW in TGM2 knockout cell line ab261876 (knockout cell lysate Human TGM2 (Transglutaminase 2) knockout A549 cell lysate ab261685). Additional cross-reactive bands were observed in the wild-type and knockout samples. Wild-type and TGM2 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. Anti-Transglutaminase 2 antibody [CUB 7402] ab2386 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4° at 1 ug/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Transglutaminase 2 antibody [CUB 7402] (Anti-Transglutaminase 2 antibody [CUB 7402] ab2386) at 1 µg/mL
Lane 1: Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 2: TGM2 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 2: Western blot - Human TGM2 (Transglutaminase 2) knockout A549 cell line (ab261876)
Lane 3: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4: HUVEC (Human umbilical vein endothelial cell line) whole cell lysate at 20 µg
Predicted band size: 77 kDa
Observed band size: 77 kDa
Western blot - Human TGM2 (Transglutaminase 2) knockout A549 cell line (ab261876)
Lanes 1 - 4: Merged signal (red and green). Green - ab64771 observed at 77 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
ab64771 was shown to specifically react with in wild-type A549 cells as signal was lost in TGM2 knockout cell line ab261876 (knockout cell lysate Human TGM2 (Transglutaminase 2) knockout A549 cell lysate ab261685). Wild-type and TGM2 knockout samples were subjected to SDS-PAGE. ab64771 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes: Anti-Transglutaminase 2 antibody (ab64771) at 1/5000 dilution
Lane 1: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: HUVEC (Human umbilical vein endothelial cell line) whole cell lysate at 20 µg
Lane 3: Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 4: TGM2 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 4: Western blot - Human TGM2 (Transglutaminase 2) knockout A549 cell line (ab261876)
Performed under reducing conditions.
Next Generation Sequencing - Human TGM2 (Transglutaminase 2) knockout A549 cell line (ab261876)
Knockout achieved by CRISPR/Cas9; X = 1 bp insertion, 8 bp deletion; Frameshift = 97%
Western blot - Human TGM2 (Transglutaminase 2) knockout A549 cell line (ab261876)
Anti-Transglutaminase 2 antibody [CUB 7402] ab2386 was shown to react with TGM2 in wild-type A549 cells in Western blot with loss of signal observed in TGM2 knockout cell line ab261876. Wild-type A549 and TGM2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with Anti-Transglutaminase 2 antibody [CUB 7402] ab2386 overnight at 4 °C at a 1/500 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.
These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.All lanes: Western blot - Anti-Transglutaminase 2 antibody [CUB 7402] (Anti-Transglutaminase 2 antibody [CUB 7402] ab2386) at 1/500 dilution
Lane 1: Wild-type A549 lysate at 40 µg
Lane 2: TGM2 knock-out A549 lysate at 40 µg
Immunocytochemistry/ Immunofluorescence - Human TGM2 (Transglutaminase 2) knockout A549 cell line (ab261876)
Anti-Transglutaminase 2 antibody [EPR28142-86] ab310333 was shown to react with TGM2 in wild-type A549 cells in immunocytochemistry with loss of signal observed in TGM2 knockout cell line ab261876. Wild-type and knockout cells were mixed and pelleted at a 1:1 ratio on coverslips. The cells were fixed with 4% paraformaldehyde (15 min) then permeabilized with 0.1% Triton X-100 (10min) and then blocked with 1x PBS, 0.01% Triton X-100, 5% BSA, 5% NGS. The cells were then incubated with Anti-Transglutaminase 2 antibody [EPR28142-86] ab310333 at 1/500 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat anti-rabbit secondary antibody to (Alexa Fluor® 555) at 0.5 ?g/ml. Acquisition of the green (wild-type), red (antibody staining) and far-red (knockout) channels was performed. Representative grayscale images of the red channel are shown. Wild-type and knockout cells are outlined with yellow and magenta dashed line, respectively. Schematic representation of the mosaic strategy used is shown on the bottom-right panel. Image was acquired with a Zeiss(LSM-880).
These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
Western blot - Human TGM2 (Transglutaminase 2) knockout A549 cell line (ab261876)
Anti-Transglutaminase 2 antibody [EPR28142-86] ab310333 was shown to react with TGM2 in wild-type A549 cells in Western blot with loss of signal observed in TGM2 knockout cell line ab261876. Wild-type A549 and TGM2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with Anti-Transglutaminase 2 antibody [EPR28142-86] ab310333 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.
These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.All lanes: Western blot - Anti-Transglutaminase 2 antibody [EPR28142-86] (Anti-Transglutaminase 2 antibody [EPR28142-86] ab310333) at 1/1000 dilution
Lane 1: Wild-type A549 lysate at 40 µg
Lane 2: TGM2 knock-out A549 lysate at 40 µg
Next Generation Sequencing - Human TGM2 (Transglutaminase 2) knockout A549 cell line (ab261876)
1 bp insertion after Glu154 (allele 1) and 8 bp deletion after Asp150 (allele 2) of the WT protein
Immunocytochemistry/ Immunofluorescence - Human TGM2 (Transglutaminase 2) knockout A549 cell line (ab261876)
Anti-Transglutaminase 2 antibody [CUB 7402] ab2386 was shown to react with TGM2 in wild-type A549 cells in immunocytochemistry with loss of signal observed in TGM2 knockout cell line ab261876. Wild-type and knockout cells were mixed and pelleted at a 1:1 ratio on coverslips. The cells were fixed with 4% paraformaldehyde (15 min) then permeabilized with 0.1% Triton X-100 (10min) and then blocked with 1x PBS, 0.01% Triton X-100, 5% BSA, 5% NGS. The cells were then incubated with Anti-Transglutaminase 2 antibody [CUB 7402] ab2386 at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat anti-rabbit secondary antibody to (Alexa Fluor® 555) at 0.5 ?g/ml. Acquisition of the green (wild-type), red (antibody staining) and far-red (knockout) channels was performed. Representative grayscale images of the red channel are shown. Wild-type and knockout cells are outlined with yellow and magenta dashed line, respectively. Schematic representation of the mosaic strategy used is shown on the bottom-right panel. Image was acquired with a Zeiss(LSM-880).
These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
Immunocytochemistry/ Immunofluorescence - Human TGM2 (Transglutaminase 2) knockout A549 cell line (ab261876)
Anti-Transglutaminase 2 antibody [EP2957] ab109200 was shown to react with TGM2 in wild-type A549 cells in immunocytochemistry with loss of signal observed in TGM2 knockout cell line ab261876. Wild-type and knockout cells were mixed and pelleted at a 1:1 ratio on coverslips. The cells were fixed with 4% paraformaldehyde (15 min) then permeabilized with 0.1% Triton X-100 (10min) and then blocked with 1x PBS, 0.01% Triton X-100, 5% BSA, 5% NGS. The cells were then incubated with Anti-Transglutaminase 2 antibody [EP2957] ab109200 at 1/300 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat anti-rabbit secondary antibody to (Alexa Fluor® 555) at 0.5 ?g/ml. Acquisition of the green (wild-type), red (antibody staining) and far-red (knockout) channels was performed. Representative grayscale images of the red channel are shown. Wild-type and knockout cells are outlined with yellow and magenta dashed line, respectively. Schematic representation of the mosaic strategy used is shown on the bottom-right panel. Image was acquired with a Zeiss(LSM-880).
These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
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