Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [5]
- Immunocytochemistry [2]
- Immunohistochemistry [1]
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Validation data
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- Product number
- MA5-25362 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- HSP40 Monoclonal Antibody (OTI1E9)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- OTI1E9
- Vial size
- 100 µL
- Concentration
- 1.00 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
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- Invitrogen Antibodies (provider)
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- Experimental details
- Western blot analysis of DNAJB1 in human tissue (1: Testis; 2: Omentum; 3: Uterus; 4: Breast; 5: Brain; 6: Liver; 7: Ovary; 8: Thyroid gland; 9: Colon) samples using 10 µg per lane. Samples were separated by SDS-PAGE and probed with DNAJB1 (Product # MA5-25362) monoclonal antibody at a dilution of 1:200.
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- Invitrogen Antibodies (provider)
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- Experimental details
- Western blot analysis of DNAJB1 in HEK293T cells in untransfected (Left lane) and transfected (Right lane) samples using 5 µg per lane. The samples were separated by SDS-PAGE and probed with DNAJB1 (Product # MA5-25362) monoclonal antibody.
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- Invitrogen Antibodies (provider)
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- Experimental details
- Western blot analysis of DNAJB1 in HeLa cells using 10 µg per lane. Samples were probed with DNAJB1 (Product # MA5-25362) monoclonal antibody at a dilution of 1:200.
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- Invitrogen Antibodies (provider)
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- Experimental details
- Knockdown of HSP40 was achieved by transfecting HeLa with HSP40 specific siRNAs (Silencer® select Product # s223882, s7009). Western blot analysis (Fig. a) was performed using whole cell extracts from the HSP40 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with HSP40 Monoclonal Antibody (OTI1E9) (Product # MA5-25362, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 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 HSP40.
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- Invitrogen Antibodies (provider)
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- Experimental details
- Western blot was performed using Anti-HSP40 Monoclonal Antibody (OTI1E9) (Product # MA5-25362) and a 40 kDa band corresponding to HSP40 was observed across cell lines tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), U-87 MG (Lane 2), MCF-7 (Lane 3), Caco-2 (Lane 4), Hep G2 (Lane 5), PC-3 (Lane 6) and A549 (Lane 7) 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 (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 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
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- Invitrogen Antibodies (provider)
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- Experimental details
- Immunofluorescent analysis of DNAJB1 in COS7 cells. Cells were transfected with a plasmid overexpressing DNAJB1 and probed with a DNAJB1 monoclonal antibody (Product # MA5-25362).
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- Invitrogen Antibodies (provider)
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- Experimental details
- Immunofluorescence analysis of HSP40 was performed using 70% confluent log phase HeLa cells and HeLa cells heat shocked at 43°C for 1 hr. 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 1 hour at room temperature. The cells were labeled with HSP40 Mouse Monoclonal Antibody (OTI1E9) (Product # MA5-25362) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then with Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32723) 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 merged image showing nuclear translocation upon heat shock. Panel e shows untreated cells with weak cytoplasmic signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
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- Invitrogen Antibodies (provider)
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- Experimental details
- Immunohistochemistry was performed on paraffin-embedded carcinoma of human bladder tissue. To expose target proteins, 10mM citric buffer, pH6.0, 100°C for 10min was used. Following antigen retrieval, tissues were probed with a DNAJB1 monoclonal antibody (Product # MA5-25362).