Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [3]
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Validation data
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- Product number
- PA5-98523 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- RAB12 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 3.25 mg/mL
- Storage
- -20°C or -80°C if preferred
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of RAB12 was achieved by transfecting HeLa with RAB12 specific siRNAs (Silencer® select Product # S223483, S47370). Western blot analysis (Fig. a) was performed using whole cell extracts from the RAB12 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with RAB12 Polyclonal Antibody (Product # PA5-98523, 1:1500) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000) and detected by chemiluminescence using the iBright™ FL1500 Imaging System (Product # A44115). 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 RAB12.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of RAB12 using a RAB12 Polyclonal antibody (Product # PA5-98523) at a concentration of 2 µg/mL. Positive WB detected in Hela whole cell lysate. A secondary Goat polyclonal antibody to rabbit IgG was applied at a 1:10,000 dilution. Observed band size: 28, 36 kDa.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using RAB12 Polyclonal Antibody (Product # PA5-98523) and a 35 kDa band corresponding to RAB12 was observed across the cell lines tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), SiHa (Lane 2), HaCaT (Lane 3), U-2 OS (Lane 4) and A549 (Lane 5) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX), 12 well. 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 (1:1500) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of RAB12 in Hela cells using a RAB12 polyclonal antibody (Product # PA5-98523) at a dilution of 1:100. Alexa Fluor 488-congugated Goat Anti-Rabbit IgG(H+L) secondary antibody was used.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of RAB12 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 45 minutes at room temperature. The cells were labeled with RAB12 Polyclonal Antibody (Product # PA5-98523, 1:100) 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) 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: Blue) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing recycling endosome membrane, lysosome membrane, golgi apparatus membrane, cytoplasmic vesicle, autophagosome and secretory localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of RAB12 in Hela cells using a RAB12 polyclonal antibody (Product # PA5-98523) at a dilution of 1:100. Alexa Fluor 488-congugated Goat Anti-Rabbit IgG(H+L) secondary antibody was used.