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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [4]
- Immunocytochemistry [2]
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- Product number
- PA5-65290 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Anti-gamma Adaptin Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- Immunogen sequence: FLLDGLSSQPL FNDIAAGIPS ITAYSKNGLK IEFTFERSNT NPSVTVITIQ ASNSTELDMT DFVFQAAVPK TFQLQLLSPS SSIVPAFNTG TI Highest antigen sequence identity to the following orthologs - mouse 98%, rat 33%.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.1 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 gamma Adaptin in human cell line RT-4, human cell line U-251 MG, human plasma, human liver tissue and human tonsil tissue. Samples were probed using a gamma Adaptin Polyclonal Antibody (Product # PA5-65290).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of gamma Adaptin in human cell line RT-4, human cell line U-251 MG, human plasma, human liver tissue and human tonsil tissue. Samples were probed using a gamma Adaptin Polyclonal Antibody (Product # PA5-65290).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of gamma Adaptin was achieved by transfecting HCT 116 with gamma Adaptin specific siRNAs (Silencer® select Product # S1142, S1143). Western blot analysis (Fig. a) was performed using Whole cell extracts from the gamma Adaptin knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with gamma Adaptin Polyclonal Antibody (Product # PA5-65290, 0.1 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knockdown confirms that antibody is specific to gamma Adaptin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-gamma Adaptin Polyclonal Antibody (Product # PA5-65290) and a 91 kDa band corresponding to gamma Adaptin was observed across cell lines and tissue extracts 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), Hep G2 (Lane 6), SH-SY5Y (Lane 7) and tissue extracts of Mouse Brain (Lane 8) and Rat Brain (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 (0.1 µg/mL) 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 Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent staining of gamma Adaptin in human cell line U-2 OS shows localization to vesicles. Samples were probed using a gamma Adaptin Polyclonal Antibody (Product # PA5-65290).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of gamma Adaptin was performed using 70% confluent log phase HCT 116 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 gamma Adaptin Polyclonal Antibody (Product # PA5-65290) at 1 µg/mL 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: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic(Golgi complex like pattern) localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
