Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [1]
- Immunohistochemistry [3]
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- Product number
- MA5-26917 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- APPL1 Monoclonal Antibody (OTI4B11)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Reactivity
- Human, Mouse, Rat
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- OTI4B11
- 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)
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- Experimental details
- Western blot analysis of APPL1 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 APPL1 (Product # MA5-26917) monoclonal antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of APPL1 was achieved by transfecting HeLa cells with APPL1 specific siRNAs (Silencer® select Product # s25038, s25039). Western blot analysis (Fig. a) was performed using whole cell extracts from the HeLa knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with APPL1 Mouse Monoclonal Antibody (Product # MA5-26917, 1:2000 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 APPL1.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-APPL1 Mouse Monoclonal Antibody (Product # MA5-26917) and 79 kDa band corresponding to APPL1 was observed across cell lines and tissues tested. Whole cell extracts (30 µg lysate) of HCT 116 (Lane 1), SW480 (Lane 2), MCF7 (Lane 3), HeLa (Lane 4), HEK-293 (Lane 5), Mouse Brain (Lane 6), Rat Brain (Lane 7), Mouse Spleen (Lane 8) and Mouse Heart (Lane 9) were electrophoresed using NuPAGE™ 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 the primary antibody (1:2000 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). A 25 kDa band (*) corresponding circulating tissue IgG was observed in mouse tissues.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of APPL1 was performed using 70% confluent log phase HeLa 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 with APPL1 Monoclonal Antibody (Product # MA5-26917) at 1:100 dilution in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32723) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green) in HeLa cells. 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). Panel d represents the merged image of HeLa cells showing cytoplasmic and 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 adenocarcinoma of human ovary tissue. To expose target proteins, heat-induced epitope retrieval by 1mM EDTA in 10mM Tris buffer (pH8.5) at 120°C for 3 min. Following antigen retrieval, tissues were probed with a APPL1 monoclonal antibody (Product # MA5-26917) at a dilution of 1:150.
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Immunohistochemistry was performed on paraffin-embedded carcinoma of human thyroid tissue. To expose target proteins, heat-induced epitope retrieval by 1mM EDTA in 10mM Tris buffer (pH8.5) at 120°C for 3 min. Following antigen retrieval, tissues were probed with a APPL1 monoclonal antibody (Product # MA5-26917) at a dilution of 1:150.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on paraffin-embedded human lymphoma tissue. To expose target proteins, heat-induced epitope retrieval by 1mM EDTA in 10mM Tris buffer (pH8.5) at 120°C for 3 min. Following antigen retrieval, tissues were probed with a APPL1 monoclonal antibody (Product # MA5-26917) at a dilution of 1:150.