Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [5]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
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Validation data
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- Product number
- MA5-25363 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- HSP40 Monoclonal Antibody (OTI1F9)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Reactivity
- Human, Mouse, Canine
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- OTI1F9
- Vial size
- 100 µL
- Concentration
- 1.6 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Western blot analysis of DNAJB1 in HepG2, HeLa, SVT2, A549, COS7, Jurkat, MDCK, PC12 and MCF7 cells using 35 µg per lane. Samples were separated by SDS-PAGE and probed with DNAJB1 (Product # MA5-25363) monoclonal antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of DNAJB1 in HepG2, HeLa, SVT2, A549, COS7, Jurkat, MDCK, PC12 and MCF7 cells using 35 µg per lane. Samples were separated by SDS-PAGE and probed with DNAJB1 (Product # MA5-25363) monoclonal antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- 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 # MA5-25363) monoclonal antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- 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 (OTI1F9) (Product # MA5-25363, 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.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-HSP40 Monoclonal Antibody (OTI1F9) (Product # MA5-25363) and a 40 kDa band corresponding to HSP40 was observed across cell lines and tissue extract tested along with an additional band at ~35 kDa. 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) and tissue extract of Mouse Testis (Lane 6) 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 SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). A non specific band around 35 kDa has also been picked up in cell lines.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- 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 # MA5-25363).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on paraffin-embedded human prostate 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 # MA5-25363).