Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [2]
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- Product number
- 711808 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- DNAJC13 Recombinant Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Other
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 0.5 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 was performed on Whole cell extracts (30 µg lysate) of COS-7 (Lane 1), U-2 OS (Lane 2), MDA-MB-231 (Lane 3), MCF-7 (Lane 4), T-47D (Lane 5), Hep G2 (Lane 6) and HEK-293 (Lane 7). The blots were probed with Anti-DNAJC13 Recombinant Rabbit Polyclonal Antibody (Product # 711808, 1 µg/mL). A 254 kDa band corresponding to DNAJC13 was observed across the cell lines tested. The blots were detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10% Bis-Tris gel (Product # NP0301BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with wet transfer method. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of RME8 was achieved by transfecting MCF7 cells with RME8 specific siRNA (Silencer® select Cat # s23548 and s23550). Western blot analysis (Fig a) was performed using whole cell extract from the RME8 knock down cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Anti-RME8 Recombinant Rabbit Polyclonal Antibody (Product # 711808, 1-3 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:5000 dilution). Densitometric analysis of this Western blot is shown in histogram (Fig b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to RME8.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- For immunofluorescence analysis, U-87 MG cells were fixed and permeabilized for detection of endogenous DNAJC13 using Anti-DNAJC13 Recombinant Rabbit Polyclonal Antibody (Product # 711808, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of DNAJC13 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating endosome localization of RME-8. 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
- Knockdown of DNAJC13 was achieved by transfecting U-87 MG cells with specific siRNA (Silencer® select Product # s23548 & s23550). Immunofluorescence analysis was performed on U-87 MG cells (untransfected, panel a-d), transfected with DNAJC13 specific siRNA (panel i-l) or non-specific scrambled siRNA (panels e-h). Cells were fixed, permeabilized, and labelled with Anti-DNAJC13 Recombinant Rabbit Polyclonal Antibody (Product # 711808, 1:100 dilution), followed by Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Nuclei (blue) were stained using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962), and Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (red) staining. Significant reduction of signal was observed upon siRNA mediated knockdown (panel i-l) confirming specificity of the antibody to DNAJC13 (green). The images were captured at 60X magnification.