Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [1]
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Validation data
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- Product number
- 711474 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- PIP5K1C Recombinant Polyclonal Antibody (16HCLC)
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is predicted to react with Monkey, Pig, Mouse, Bovine
- Antibody clone number
- 16HCLC
- Concentration
- 0.5 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on Whole cell extracts (30 µg lysate) of A-431 (Lane 1), HEK-293 (Lane 2), HeLa (Lane 3), T-47D (Lane 4) and A549 (Lane 5). The blots were probed with Anti-Phosphatidylinositol 4-phosphate 5-kinase type-1 gamma (PIP5K1C) Recombinant Rabbit Polyclonal Antibody (Product # 711474, 2.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:4000 dilution). A ~86 kDa band corresponding to Phosphatidylinositol 4-phosphate 5-kinase type-1 gamma (PIP5K1C) was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on Whole cell extracts (30 µg lysate) of A-431 (Lane 1), HEK-293 (Lane 2), HeLa (Lane 3), T-47D (Lane 4) and A549 (Lane 5). The blots were probed with Anti-Phosphatidylinositol 4-phosphate 5-kinase type-1 gamma (PIP5K1C) Recombinant Rabbit Polyclonal Antibody (Product # 711474, 2.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:4000 dilution). A ~86 kDa band corresponding to Phosphatidylinositol 4-phosphate 5-kinase type-1 gamma (PIP5K1C) was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- For immunofluorescence analysis, A431 cells were fixed and permeabilized for detection of endogenous Phosphatidylinositol 4-phosphate 5-kinase type-1 gamma using Anti- PIP5K1C Recombinant Rabbit Polyclonal Antibody (Product # 711474, 5 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of Phosphatidylinositol 4-phosphate 5-kinase type-1 gamma protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating cytoplasmic and nuclear localisation of PIP5K1C. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.