Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [1]
Submit
Validation data
Reference
Comment
Report error
- Product number
- PA1-16604 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- PINK1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 1 mg/mL
- Storage
- 4° C, do not freeze
No comments: Submit comment
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot detection of murine PINK1 using Product # PA1-16604. 1: molecular weight marker. 2. MES cell Mitochondria (20 µg). 3. MES cytosol (20 µg). 4. MES nuclear (20 µg) as negative control. 5. Purified human cytochrome C (0.1 µg) as PINK1 negative control.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of PINK1 in 20 µg MES cell Mitochondria, MES cytosol, MES nuclear. Samples were incubated in PINK1 polyclonal antibody (Product # PA1-16604). Lane 1: molecular weight marker; Lane 2: MES cell Mitochondria with a band at the observed molecular weight of 63 kDa; Lane 3: MES cytosol; Lane 4: MES nuclear as negative control; Lane 5: Purified human cytochrome C (0.1 µg) as PINK1 negative control.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of PINK1 was performed by loading 30 µg of U-2 OS (Lane 1), U-2 OS - PINK1 knockout (Lane 2) Whole cell extracts. The blot was probed with Anti-PINK1 Polyclonal Antibody (Product # PA1-16604,1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Loss of signal upon CRISPR mediated knockout (KO) confirms that antibody is specific to PINK1.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-PINK1 Polyclonal Antibody (Product # PA1-16604) and a 63kDa band corresponding to PINK1 was observed across the cell lines tested. Whole cell extracts (30 µg lysate) of U-2 OS (Lane 1), HeLa (Lane 2), SH-SY5Y (Lane 3) and U-87 MG (Lane 4) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody at 1:1000 dilution and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of PINK1 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 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 PINK1 Polyclonal Antibody (Product # PA1-16604) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034), (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 dilution). Panel d represents the merged image showing Mitochondrial localization. Panel e represents control HeLa cells with no primary antibody to assess background. The images were captured at 60X magnification.