Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [2]
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Validation data
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- Product number
- PA5-67357 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- C2orf33 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- Immunogen sequence: DLIQSTPFKPL ALKTPPRVLT LSERPLDFLD LERPPTTPQN EEIRAVGRLK RERSMSENAV RQNGQLVRND SLWH Highest antigen sequence identity to the following orthologs - mouse 93%, rat 96%.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.2 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
- KD of C2orf33 was achieved by transfecting HeLa cells with C2orf33 specific siRNAs (Silencer® select Product # s25664, s25665). Western blot analysis (Fig. a) was performed using Membrane enriched extracts from the C2orf33 KD cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with C2orf33 Polyclonal Antibody (Product # PA5-67357, 1:1000 dilution) and Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (Product # A27036, 1:4000 dilution). 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 C2orf33.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-C2orf33 Rabbit Polyclonal Antibody (Product # PA5-67357) and 37kDa, 36kDa, 28kDa, 27kDa bands corresponding to isoforms of C2orf33 were observed across cell lines and tissues tested. Membrane enriched extracts (30 µg lysate) of HeLa (Lane 1), Hep G2 (Lane 2), A549 (Lane 3), KARPAS 299 (Lane 4), DU 145 (Lane 5), LNCaP (Lane 6), SH-SY5Y (Lane 7), A-431 (Lane 8), tissue extracts of Mouse Heart (Lane 9) and Mouse Brain (Lane 10) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris 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 (1:1000 dilution) and detected by Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (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 C2orf33 in human cell line SiHa shows localization to mitochondria. Samples were probed using a C2orf33 Polyclonal Antibody (Product # PA5-67357).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of C2orf33 was performed using 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 1% BSA for 1 hour at room temperature. The cells were labeled with C2orf33 Rabbit Polyclonal Antibody (Product # PA5-67357) at 4µg/mL dilution in 0.1% BSA and incubated overnight at 4 degree 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 composite image showing mitochondrial localization of C2orf33. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification. .