Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [1]
- Immunocytochemistry [1]
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Validation data
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- Product number
- LF-MA0212 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Anti-SHP-1 Monoclonal Antibody (14D5)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Description
- A suggested positive control for this product is HL-60 cells.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 14D5
- Vial size
- 100 µL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on nuclear enriched extracts (30 µg lysate) of Raji (Lane 1), Jurkat (Lane 2), MCF7 (Lane 3) and Caco-2 (Lane 4). The blot was probed with Anti-SHP-1 Mouse Monoclonal Antibody (Product # LF-MA0212, 1:2,000 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A28177, 0.4 µg/mL, 1:2500 dilution). A 67 kDa band corresponding to SHP-1 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of SHP-1 Monoclonal Antibody was performed using 70% confluent log phase K562 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with SHP-1 (14D5) Mouse Monoclonal Antibody (Product # LF-MA0212) at 1:250 dilution in 0.1% BSA, incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (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 cytoplasmic and membranous localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification