Antibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [1]
- Immunohistochemistry [3]
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Validation data
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- Product number
- MA5-26346 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- GCLC Monoclonal Antibody (OTI1A3)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Reactivity
- Human, Mouse, Rat
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- OTI1A3
- Vial size
- 100 µL
- Concentration
- 1 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
Submitted references Spatial locations of certain enzymes and transporters within preinvasive ductal epithelial cells predict human breast cancer recurrences.
Kraft AM, Petty HR
American journal of physiology. Cell physiology 2020 Nov 1;319(5):C910-C921
American journal of physiology. Cell physiology 2020 Nov 1;319(5):C910-C921
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of GCLC in HEK293T cells in untransfected (Left lane) and transfected (Right lane) samples using 5 µg per lane. The samples were separated by SDS-PAGE and probed with GCLC (Product # MA5-26346) monoclonal antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of GCLC in HepG2, HeLa, SVT2, A549, COS7, Jurkat, MDCK, PC12 and MCF7 cells using 35 µg per lane. Samples were separated by SDS-PAGE and probed with GCLC (Product # MA5-26346) monoclonal antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of GCLC was achieved by transfecting Hep G2 cells with GCLC specific siRNAs (Silencer® select Product # s5800). Western blot analysis (Fig. a) was performed whole cell extracts from the GCLC knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with GCLC Monoclonal Antibody (Product # MA5-26346, 1:500 dilution) and Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 0.25 µg/ml, 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 GCLC.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-GCLC Monoclonal Antibody (Product # MA5-26346) and a 72kDa band corresponding to GCLC was observed across cell lines and tissues tested. Whole cell extracts (30 ug lysate) of A549 (Lane 1), Hep G2 (Lane 2), MCF7 (Lane 3), Jurkat (Lane 4), Caco-2 (Lane 5), Mouse Liver (Lane 6), Mouse Kidney (Lane 7) and Mouse Stomach (Lane 8) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris 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 the primary antibody (1:500 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 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
- Immunofluorescence analysis GCLC Monoclonal Antibody (OTI1A3) was performed using 70% confluent log phase HepG2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled GCLC monoclonal antibody (Product # MA5-26346) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e 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
- Immunohistochemistry was performed on paraffin-embedded human pancreas tissue. To expose target proteins, 1 mM EDTA in 10mM Tris, pH8.5, 120°C for 3min was used. Following antigen retrieval, tissues were probed with a GCLC monoclonal antibody (Product # MA5-26346).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on paraffin-embedded carcinoma of human thyroid tissue. To expose target proteins, 1 mM EDTA in 10mM Tris, pH8.5, 120°C for 3min was used. Following antigen retrieval, tissues were probed with a GCLC monoclonal antibody (Product # MA5-26346).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on paraffin-embedded human lymph node tissue. To expose target proteins, 1 mM EDTA in 10mM Tris, pH8.5, 120°C for 3min was used. Following antigen retrieval, tissues were probed with a GCLC monoclonal antibody (Product # MA5-26346).