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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [4]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
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- Product number
- PA5-54146 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- PPA1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- Immunogen sequence: NAEFKDKDFA IDIIKSTHDH WKALVTKKTN GKGISCMNTT LSESPFKCDP DAARAIVDAL PPPCESACTV PTDVDKWFHH Q
- Concentration
- 0.05 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-PPA1 Rabbit Polyclonal Antibody (Product # PA5-54146) and 32 kDa band corresponding to PPA1 was observed across samples tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), U-87 MG (Lane 2), HT-29 (Lane 3), HCT 116 (Lane 4), SW-480 (Lane 5), MCF-7 (Lane 6) and tissue extract of Mouse Small Intestine (Lane 7) were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris 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 (0.4µg/mL) and detected by Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of PPA1 using PPA1 Polyclonal Antibody (Product # PA5-54146) (A) shows similar pattern to an independent PPA1 Polyclonal Antibody (B).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- KD of PPA1 was achieved by transfecting U-87 MG with PPA1 specific siRNAs (Silencer® select Product # s10878, s10877). Western blot analysis (Fig. a) was performed using whole cell extracts from the PPA1 KD cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with PPA1 Polyclonal Antibody (Product # PA5-54146, 0.4µg/mL) and Goat Anti Rabbit (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 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 PPA1.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-PPA1 Rabbit Polyclonal Antibody (Product # PA5-54146) and 32 kDa band corresponding to PPA1 was observed across samples tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), U-87 MG (Lane 2), HT-29 (Lane 3), HCT 116 (Lane 4), SW-480 (Lane 5), MCF-7 (Lane 6) and tissue extract of Mouse Small Intestine (Lane 7) were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris 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 (0.4µg/mL) and detected by Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (Product # A27036, 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
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent staining of PPA1 in human cell line U-2 OS shows positivity in vesicles. Samples were probed using a PPA1 Polyclonal Antibody (Product # PA5-54146).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical staining of PPA1 in human testis using a PPA1 Polyclonal Antibody (Product # PA5-54146) shows cytoplasmic and nuclear positivity in cells in seminiferous ducts and Leydig cells.
