Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [2]
- Immunohistochemistry [1]
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Validation data
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- Product number
- PA5-76618 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Syntenin 1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen and the purity is > 95% (by SDS-PAGE).
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 1 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 Syntenin 1. Samples were incubated with Syntenin 1 polyclonal antibody (Product # PA5-76618).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Syntenin 1 Polyclonal Antibody (Product # PA5-76618) and a 32kDa band corresponding to Syntenin-1 was observed across cell lines tested. Whole cell extracts (30 µg lysate) of MDA-MB-231 (Lane 1), T-47D (Lane 2), MCF7 (Lane 3), HT-29 (Lane 4), HCT 116 (Lane 5), U-87 MG (Lane 6), DU 145 (Lane 7), HeLa (Lane 8) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0301BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23002) 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, 1:6000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). Syntenin 1 is overexpressed and promotes cell migration in metastatic human breast cancer cell lines (doi: 10.1038/sj.onc.1205514). Hence, the expression was observed to be high in MDA-MB-231 when compared to MCF7 and T-47D. In addition to this, the expression level was found to be high in HT-29 when compared to HCT 116. An uncharacterized band was observed at ~35kDa.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Syntenin 1 in U2OS cells. Samples were incubated with Syntenin 1 polyclonal antibody (Product # PA5-76618).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Syntenin-1 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 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Syntenin 1 Polyclonal Antibody (Product # PA5-76618) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # 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). Panel d represents the merged image showing nuclear and cytoplasmic localization. Panel e represents MCF7 cells with lower expression level of Syntenin 1 when compared to MDA-MB-231. 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
- Immunohistochemistry analysis of Syntenin 1 in paraffin-embedded human stomach tissue. Samples were incubated with Syntenin 1 polyclonal antibody (Product # PA5-76618).