Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [1]
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Validation data
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- Product number
- PA5-17820 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- alpha Synuclein Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- It is not recommended to aliquot this antibody.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 60 µg/mL
- Storage
- -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of alpha Synuclein was performed by loading 30 µg of HeLa control (Lane 1), HeLa - alpha Synuclein knockout (Lane 2) whole cell lysates. The blot was probed with alpha Synuclein Polyclonal Antibody (Product # PA5-17820, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL 1:4000 dilution). Loss of signal upon CRISPR mediated knockout (KO) confirms that antibody is specific to alpha Synuclein. An uncharacterized band was observed at ~32 kDa in the samples tested.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on tissue extracts (30 µg lysate) of Mouse Brain (Lane 1), Rat Pup Brain (Lane 2) and Mouse Liver (Lane 3). The blot was probed with Anti-alpha Synuclein Polyclonal Antibody (Product # PA5-17820, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 15 kDa band corresponding to alpha Synuclein was detected in Mouse Brain, Rat Pup Brain and not in Mouse Liver which is reported negative for alpha Synuclein expression. An uncharacterized band was observed at ~32 kDa in all the tissues tested.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of alpha synuclein was performed using 70% confluent log phase SH-SY5Y cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with alpha Synuclein Polyclonal Antibody (Product # PA5-17820) at 1:250 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 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 cytosolic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.