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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [5]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
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- Product number
- PA5-11412 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- HTRA1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human, Mouse
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 200 µL
- Concentration
- 0.5 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of HtrA1 using a HtrA1 polyclonal antibody (Product # PA5-11412) mouse brain tissue lysate.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using HTRA1 Polyclonal Antibody (Product # PA5-11412) and a 51 kDa band corresponding to HTRA1 was observed in SK-OV-3, MCF 10A, PC-3, HT-1080 and BJ but not in MCF7 and LNCaP which are low expressing cell lines (10.1371/journal.pone.0039446, 10.7150/jca.5702). Whole cell extracts (50 µg lysate) of SK-OV-3 (Lane 1), MCF10A (Lane 2), MCF7 (Lane 3), PC-3 (Lane 4), LNCaP (Lane 5), HT-1080 (Lane 6) and BJ (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23002) by iBlot® 2 Gel Transfer Device (Product # IB21001). The blot was probed with the primary antibody (1:2000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain) 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).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using HTRA1 Polyclonal Antibody (Product # PA5-11412) and a 51 kDa band corresponding to HTRA1 was observed in conditioned media from MCF 10A but not in MCF7. Conditioned media (10 ul) of MCF 10A (Lane 1) and MCF7 (Lane 2) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23002) by iBlot® 2 Gel Transfer Device (Product # IB21001). The blot was probed with the primary antibody (1:2000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain) 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).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of HTRA1 was achieved by transfecting transfecting MCF 10A cells with HTRA1 specific siRNAs (Silencer® select Product # S11281, S533018). Western blot analysis (Fig. a) was performed using whole cell extracts from the HTRA1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with HTRA1 Polyclonal Antibody (Product # PA5-11412, 1:2000 dilution) and Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000). 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 HTRA1.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of HTRA1 was achieved by transfecting MCF 10A cells with HTRA1 specific siRNAs (Silencer® select Product # S11281, S533018). Western blot analysis was performed using conditioned media from HTRA1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The conditioned media was quantified using the Pierce™ Rapid Gold BCA Protein Assay Kit (Product # A53225) and normalized before loading. The blot was probed with HTRA1 Polyclonal Antibody (Product # PA5-11412, 1:2000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000 dilution). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to HTRA1.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of HeLa cells using a HtrA1 polyclonal antibody (Product # PA5-11412) at a dilution of 1:10-50, followed by a fluor-conjugated goat anti-rabbit secondary antibody (green). Nuclei were stained with DAPI (blue).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis in formalin-fixed, paraffin-embedded human kidney tissue using a HtrA1 polyclonal antibody (Product # PA5-11412), followed by HRP-conjugated secondary antibody and DAB staining.
