Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [1]
- Immunohistochemistry [8]
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Validation data
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- Product number
- MA5-25076 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- SHC Monoclonal Antibody (OTI3A1)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Reactivity
- Human, Mouse, Rat
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- OTI3A1
- Vial size
- 100 µL
- Concentration
- 1 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunoprecipitation was performed on human HBL-100/MCF-7 cell lysate using 1 µg of protein. Antigen-antibody complexes were formed by incubating 2 µg of sample with SHC1 monoclonal antibody (Product # MA5-25076) antibody, and visualizing the result using Super Signal West Femto chemiluminescence substrate.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of SHC1 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 SHC1 (Product # MA5-25076) monoclonal antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of SHC was achieved by transfecting A-431 with SHC specific siRNAs (Silencer® select Product # S12813, S12812). Western blot analysis (Fig. a) was performed using whole cell extracts from the SHC knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with SHC Monoclonal Antibody (OTI3A1) (Product # MA5-25076, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 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 SHC.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-SHC Monoclonal Antibody (OTI3A1) (Product # MA5-25076) and 46 and 52kDa bands corresponding to SHC were observed along with an uncharacterized band (*) at ~ 37kDa across all cell lines tested. Whole cell extracts (30µg lysate) of A-431 (Lane 1), HEK-93 (Lane 2), HeLa (Lane 3), Jurkat (Lane 4), MCF7 (Lane 5) and NIH/3T3 (Lane 6) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). 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-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 SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of SHC1 in COS7 cells. Cells were transfected with a plasmid overexpressing SHC1 and probed with a SHC1 monoclonal antibody (Product # MA5-25076).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Immunohistochemistry was performed on paraffin-embedded breast tissue. To expose target proteins, 10mM citric buffer, pH6.0, 100°C for 10min was used. Following antigen retrieval, tissues were probed with a SHC1 monoclonal antibody (Product # MA5-25076) at a dilution of 1:50.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on paraffin-embedded colon tissue. To expose target proteins, 10mM citric buffer, pH6.0, 100°C for 10min was used. Following antigen retrieval, tissues were probed with a SHC1 monoclonal antibody (Product # MA5-25076) at a dilution of 1:50.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on paraffin-embedded kidney tissue. To expose target proteins, 10mM citric buffer, pH6.0, 100°C for 10min was used. Following antigen retrieval, tissues were probed with a SHC1 monoclonal antibody (Product # MA5-25076) at a dilution of 1:50.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on paraffin-embedded carcinoma of liver tissue. To expose target proteins, 10mM citric buffer, pH6.0, 100°C for 10min was used. Following antigen retrieval, tissues were probed with a SHC1 monoclonal antibody (Product # MA5-25076) at a dilution of 1:50.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on paraffin-embedded lung tissue. To expose target proteins, 10mM citric buffer, pH6.0, 100°C for 10min was used. Following antigen retrieval, tissues were probed with a SHC1 monoclonal antibody (Product # MA5-25076) at a dilution of 1:50.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on paraffin-embedded prostate tissue. To expose target proteins, 10mM citric buffer, pH6.0, 100°C for 10min was used. Following antigen retrieval, tissues were probed with a SHC1 monoclonal antibody (Product # MA5-25076) at a dilution of 1:50.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on paraffin-embedded carcinoma of prostate tissue. To expose target proteins, 10mM citric buffer, pH6.0, 100°C for 10min was used. Following antigen retrieval, tissues were probed with a SHC1 monoclonal antibody (Product # MA5-25076) at a dilution of 1:50.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on paraffin-embedded bladder tissue. To expose target proteins, 10mM citric buffer, pH6.0, 100°C for 10min was used. Following antigen retrieval, tissues were probed with a SHC1 monoclonal antibody (Product # MA5-25076) at a dilution of 1:50.