Human TGM2 (Transglutaminase 2) knockout A549 cell line
- Advanced Validation
- What is this?
Be the first to review this product! Submit a review
|
(0 Publication)
TGM2 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9 X = 1 bp insertion 8 bp deletion Frameshift = 97%. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View 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
- ICC/IF
Lab
Immunocytochemistry/ Immunofluorescence - Human TGM2 (Transglutaminase 2) knockout A549 cell line (AB261876)
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 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 µg/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.
- ICC/IF
Lab
Immunocytochemistry/ Immunofluorescence - Human TGM2 (Transglutaminase 2) knockout A549 cell line (AB261876)
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 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 µg/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.
- ICC/IF
Lab
Immunocytochemistry/ Immunofluorescence - Human TGM2 (Transglutaminase 2) knockout A549 cell line (AB261876)
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 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 µg/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.
- WB
Lab
Western blot - Human TGM2 (Transglutaminase 2) knockout A549 cell line (AB261876)
Lanes 1 - 4 : Merged signal (red and green). Green - ab2386 observed at 77 kDa. Red - loading control ab181602 observed at 37 kDa.
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 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. ab2386 and 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 ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed 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] (<a href='/en-us/products/primary-antibodies/transglutaminase-2-antibody-cub-7402-ab2386'>ab2386</a>) 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
false
- WB
Lab
Western blot - Human TGM2 (Transglutaminase 2) knockout A549 cell line (AB261876)
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 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] (<a href='/en-us/products/primary-antibodies/transglutaminase-2-antibody-cub-7402-ab2386'>ab2386</a>) at 1/500 dilution
Lane 1:
Wild-type A549 lysate at 40 µg
Lane 2:
TGM2 knock-out A549 lysate at 40 µg
false
- WB
Lab
Western blot - Human TGM2 (Transglutaminase 2) knockout A549 cell line (AB261876)
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 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] (<a href='/en-us/products/primary-antibodies/transglutaminase-2-antibody-epr28142-86-ab310333'>ab310333</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 lysate at 40 µg
Lane 2:
TGM2 knock-out A549 lysate at 40 µg
false
- WB
Lab
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, 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 ab261685). Wild-type and TGM2 knockout samples were subjected to SDS-PAGE. ab64771 and 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 ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed 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)
false
- NGS
Supplier Data
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%
- NGS
Supplier Data
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
- WB
Lab
Western blot - Human TGM2 (Transglutaminase 2) knockout A549 cell line (AB261876)
This data was developed using ab109200, the same antibody clone in a different buffer formulation.
ab109200 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 ab109200 overnight at 4 °C at a 1/10000 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 [EP2957] (<a href='/en-us/products/primary-antibodies/transglutaminase-2-antibody-ep2957-ab109200'>ab109200</a>) at 1/10000 dilution
Lane 1:
Wild-type A549 lysate at 40 µg
Lane 2:
Western blot - Human TGM2 (Transglutaminase 2) knockout A549 cell line (ab261876) at 40 µg
Observed band size: 75 kDa
false
Reactivity data
Product details
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Complementary Products
Primary Antibodies
AB109200
Anti-Transglutaminase 2 antibody [EP2957]
KO Validated|RabMAb|Recombinant
![Immunocytochemistry/ Immunofluorescence - Anti-Transglutaminase 2 antibody [EP2957] (AB109200)](https://content.abcam.com/products/images/transglutaminase-2-antibody-ep2957-ab109200--immunocytochemistry-immunofluorescence-img438567.jpg)
- 4
- 1
Cell Lines & Lysates
AB255343
Human SQSTM1 (p62) knockout HEK-293T cell line
Advanced Validation
- 0
- 0
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com