Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [5]
- Immunocytochemistry [2]
- Immunohistochemistry [2]
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- Product number
- PA5-29357 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- TGM2 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: H1299, HeLa, HepG2. Predicted reactivity: Mouse (84%), Rat (82%), Bovine (88%), Guinea pig (82%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
- Reactivity
- Human, Mouse
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 1.05 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Transglutaminase 2 using 30 µg of HepG2 lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a Transglutaminase 2 polyclonal antibody (Product # PA5-29357) at a dilution of 1:1000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockout of TGM2 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR979226_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of TGM2 was performed by loading 30 µg of HeLa Wild Type (Lane 1), HeLa Cas9 (Lane 2) andHela TGM2 KO (Lane 3) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-TGM2 Polyclonal Antibody (Product # PA5-29357, 1:1,000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:6,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to TGM2.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using TGM2 Polyclonal Antibody (Product # PA5-29357). Various whole cell extracts (30 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with TGM2 Polyclonal Antibody (Product # PA5-29357) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of TGM2 was achieved by transfecting HeLa with TGM2 specific siRNAs (Silencer® select Product # S14087, S14088). Western blot analysis (Fig. a) was performed using Whole cell extracts from the TGM2 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with TGM2 Polyclonal Antibody (Product # PA5-29357, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to TGM2.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-TGM2 Polyclonal Antibody(Product # PA5-29357) and a 78kDa band corresponding to TGM2 was observed across cell lines and tissues tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), A549 (Lane 2), MDA-MB-231 (Lane 3), MCF7 (Lane 4), LNCaP (Lane 5) and DU 145 (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry-Immunofluorescence analysis of TGM2 was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: TGM2 Polyclonal Antibody (Product # PA5-29357) diluted at 1:500. Red: alpha Tubulin, a cytoskeleton marker. Blue: Hoechst 33342 staining.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of TGM2 was performed using 70% confluent log phase MDA-MB-231 and MCF7 cells. The cells were fixed with 4% Paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 10 minutes at room temperature. The cells were labeled with TGM2 Polyclonal Antibody (Product # PA5-29357) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Alexa Fluor Plus 488 donkey anti-rabbit IgG secondary antibody -A32790), (1:2000 dilution) for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing cytoplasmic localization. Panel e represents MCF7 cells having no expression of TGM2. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of paraffin-embedded H441 xenograft, using Transglutaminase 2 (Product # PA5-29357) antibody at 1:500 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- TGM2 Polyclonal Antibody detects Transglutaminase 2 protein at cytoplasm on mouse heart by immunohistochemical analysis. Sample: Paraffin-embedded mouse heart. TGM2 Polyclonal Antibody (Product # PA5-29357) diluted at 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.