702444

antibody from Invitrogen Antibodies

Targeting: PIP5K1C KIAA0589, LCCS3, PIP5Kgamma

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

702444

Provider

Invitrogen Antibodies

Product name

PIP5K1C Recombinant Rabbit Monoclonal Antibody (16H4L5)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with Monkey, Pig, Mouse, Bovine

Antibody clone number

16H4L5

Concentration

0.5 mg/mL

References

Submitted references

Lipid Messenger Phosphatidylinositol-4,5-Bisphosphate Is Increased by Both PPARα Activators and Inhibitors: Relevance for Intestinal Cell Differentiation.

Cizkova K, Koubova K, Tauber Z
Biology 2022 Jun 30;11(7)

Western blot

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), HeLa (Lane 2) and A549 (Lane 3). The blots were probed with Anti-Phosphatidylinositol 4-phosphate 5-kinase type-1 gamma (PIP5K1C) Recombinant Rabbit Monoclonal Antibody (Product # 702444, 2.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 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).

Immunocytochemistry

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 Monoclonal Antibody (Product # 702444, 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 nuclear localisation of PIP5K1C. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Relationships between levels of PPARalpha and PIP5K1C, PIP2, PI3K, and PTEN in HT-29 cell line. ( A ) Spearman correlation coefficients ( n = 12) for expression levels obtained with In-Cell ELISA method. PPARalpha expression under the same experimental condition was examined in our previous study []. Moderate positive correlations were found between PPARalpha and PIP5K1C and PPARalpha and PIP2, whereas PPARalpha and PTEN expression showed moderate negative correlation (statistical significance marked with asterisk, * p