Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [7]
- Immunohistochemistry [6]
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- Product number
- MA5-32482 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- PKC delta Recombinant Rabbit Monoclonal Antibody (JJ093-02)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Description
- Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- JJ093-02
- Vial size
- 100 µL
- Concentration
- 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)
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- Experimental details
- Western blot analysis of PKC delta in different lysates using a Monoclonal antibody (Product #MA5-32482) at a dilution of 1:1,000. Positive control: Lane 1: Hela, Lane 2: NIH/3T3.
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- Invitrogen Antibodies (provider)
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- Experimental details
- Knockdown of PRKCD was achieved by transfecting THP-1 with PRKCD specific siRNAs (Silencer® select Product # S11099). Western blot analysis (Fig. a) was performed using whole cell extracts from the PRKCD knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with PKC delta Recombinant Rabbit Monoclonal Antibody (JJ093-02) (Product # MA5-32482, 1:2000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000 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 PRKCD.
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- Invitrogen Antibodies (provider)
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- Experimental details
- Western blot was performed using PKC delta Recombinant Rabbit Monoclonal Antibody (JJ093-02) (Product # MA5-32482) and a 78 kDa band corresponding to PRKCD was observed across cell lines and tissue tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), BeWo (Lane 2), Raji (Lane 3), THP-1 (Lane 4), NIH/3T3 (Lane 5), MCF7 (Lane 6), BT-549 (Lane 7), HCT 116 (Lane 8), tissue extract of Rat Brain (Lane 9) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX), 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 (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580). Expression of PRKCD was observed to be high in MCF-7 in comparison to BT-549 (Ref :https://doi.org/10.3389/fonc.2019.01323).
Supportive validation
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- Invitrogen Antibodies (provider)
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- Experimental details
- Immunocytochemical analysis of PKC delta in A549 cells using a PKC delta Monoclonal antibody (Product # MA5-32482) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
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- Invitrogen Antibodies (provider)
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- Experimental details
- Immunocytochemical analysis of PKC delta in MCF-7 cells using a PKC delta Monoclonal antibody (Product # MA5-32482) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
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- Invitrogen Antibodies (provider)
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- Experimental details
- Immunocytochemical analysis of PKC delta in HepG2 cells using a PKC delta Monoclonal antibody (Product # MA5-32482) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
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- Invitrogen Antibodies (provider)
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- Experimental details
- Immunocytochemical analysis of PKC delta in RH-35 cells using a PKC delta Monoclonal antibody (Product # MA5-32482) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of PRKCD was performed using 70% confluent log phase HeLa treated with Etoposide (100 µM for 1hr). 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 PKC delta Recombinant Rabbit Monoclonal Antibody (JJ093-02) (Product # MA5-32482) at 1:100 dilution 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 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 predominant nuclear nuclear as well as faint cytoplasmic localization. Panel e represents less expression of PKC delta. Panel f 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
- Immunofluorescence analysis of PRKCD was performed using 70% confluent log phase BeWo 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 PKC delta Recombinant Rabbit Monoclonal Antibody (JJ093-02) (Product # MA5-32482) at 1:100 dilution 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 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with Hoechst 33342 (Product # H1399). F-actin (Panel c: Red) was stained with Alexa Fluor™ Plus 647 Phalloidin (Product # A30107, 1:2000 dilution). Panel d represents the merged image showing nuclear as well as cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 40X magnification in CellInsight CX7 LZR High-Content Screening (HCS) Platform (Product # CX7A1110LZR) and externally deconvoluted (D.Sage et al./Methods 115 (2017) 28–41).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of PRKCD was performed using 70% confluent log phase HEK 293 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 PKC delta Recombinant Rabbit Monoclonal Antibody (JJ093-02) (Product # MA5-32482) at 1:100 dilution 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 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with Hoechst 33342 (Product # H1399). F-actin (Panel c: Red) was stained with Alexa Fluor™ Plus 647 Phalloidin (Product # A30107, 1:2000 dilution). Panel d represents the merged image showing nuclear as well as cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 40X magnification in CellInsight CX7 LZR High-Content Screening (HCS) Platform (Product # CX7A1110LZR) and externally deconvoluted (D.Sage et al./Methods 115 (2017) 28–41).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Immunohistochemical analysis of PKC delta of paraffin-embedded Mouse testis tissue using a PKC-delta Monoclonal antibody (Product #MA5-32482). Counter stained with hematoxylin.
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- Experimental details
- Immunohistochemical analysis of PKC delta of paraffin-embedded Mouse liver tissue using a PKC-delta Monoclonal antibody (Product #MA5-32482). Counter stained with hematoxylin..
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- Immunohistochemical analysis of PKC delta of paraffin-embedded Human tonsil tissue using a PKC-delta Monoclonal antibody (Product #MA5-32482). Counter stained with hematoxylin.
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- Experimental details
- Immunohistochemical analysis of PKC delta of paraffin-embedded Human liver tissue using a PKC-delta Monoclonal antibody (Product #MA5-32482). Counter stained with hematoxylin.
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- Experimental details
- Immunohistochemical analysis of PKC delta of paraffin-embedded Human spleen tissue using a PKC-delta Monoclonal antibody (Product #MA5-32482). Counter stained with hematoxylin.
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- Invitrogen Antibodies (provider)
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- Experimental details
- Immunohistochemical analysis of PKC delta of paraffin-embedded Human breast carcinoma tissue using a PKC-delta Monoclonal antibody (Product #MA5-32482). Counter stained with hematoxylin.