Revision 3

#61407Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IHC-P, IF-F, IF-IC, FC-FP

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

50

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q8TBX8

Entrez-Gene Id:

79837

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunohistochemistry (Paraffin) 1:800 - 1:3200
Immunofluorescence (Frozen) 1:50 - 1:200
Immunofluorescence (Immunocytochemistry) 1:200 - 1:800
Flow Cytometry (Fixed/Permeabilized) 1:50 - 1:200

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

PIP4K2C (E7P3S) Rabbit mAb recognizes endogenous levels of total PIP4K2C protein. This antibody does not cross-react with PIP4K2A or PIP3K2B proteins.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu321 of human PIP4K2C protein.

Background

Phosphatidylinositol-5-phosphate 4-kinases (PIP4Ks) synthesize phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P2), a key precursor in phosphoinositide signaling that directly modulates the activity of signaling proteins and cellular processes. There are two subfamilies of PIP kinases, type I and II, that generate PtdIns(4,5)P2 from distinct substrate pools. PIP4 type I kinases use PtdIns5P as a substrate, whereas PIP5 type II kinases use PtdIns4P (1,2). In mammalian cells, three isoforms of each PIP4K and PIP5K subfamily, encoded by distinct genes, have been characterized (3-7). All PIP kinases are stimulated by phosphatidic acid, extensively regulated by ARF and Rho GTPases, and inhibited by protein kinase A and PI-stimulated autophosphorylation (8).

PIP4K2C, also known as phosphatidylinositol-5-phosphate 4-kinase type-2 gamma (PI5P4Kγ), is a member of the type II PIP kinase family, and is broadly expressed with the highest levels in kidney and brain tissue (9). PIP4K2C can form a homodimer or heterodimer with its two other known family members, PIP4K2A and PIP4K2B (10). PIP4K2C is a regulator of mammalian target of rapamycin complex 1 (mTORC1) signaling, and Pip4k2c KO mice demonstrate enhanced immune responses and develop autoimmunity (11,12).

  1. Rameh, L.E. et al. (1997) Nature 390, 192-6.
  2. Zhang, X. et al. (1997) J Biol Chem 272, 17756-61.
  3. Ishihara, H. et al. (1996) J Biol Chem 271, 23611-4.
  4. Loijens, J.C. and Anderson, R.A. (1996) J Biol Chem 271, 32937-43.
  5. Ishihara, H. et al. (1998) J Biol Chem 273, 8741-8.
  6. Itoh, T. et al (1998) J. Biol. Chem. 273, 20292-20299
  7. Boronenkov, I.V. et al. (1998) Mol Biol Cell 9, 3547-60.
  8. Oude Weernink, P.A. et al. (2004) Eur J Pharmacol 500, 87-99.
  9. Clarke, J.H. et al. (2008) Am J Physiol Renal Physiol 295, F1422-30.
  10. Ciruela, A. et al. (2000) Biochem J 346 Pt 3, 587-91.
  11. Mackey, A.M. et al. (2014) Sci Signal 7, ra104.
  12. Shim, H. et al. (2016) Proc Natl Acad Sci USA 113, 7596-601.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

使用に関する制限

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Revision 3
#61407

PIP4K2C (E7P3S) Rabbit mAb

Western Blotting Image 1: PIP4K2C (E7P3S) Rabbit mAb Expand Image
Western blot analysis of extracts from various human, mouse, and rat cell lines using PIP4K2C (E7P3S) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). Negative expression of PIP4K2C protein in TALL-1 and REH cells is consistent with the predicted expression pattern.
Western Blotting Image 2: PIP4K2C (E7P3S) Rabbit mAb Expand Image
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected (+) with constructs expressing HA-tagged full-length human PIP4K2A (hPIP4K2A-HA), HA-tagged full-length human PIP4K2B (hPIP4K2B-HA), or Myc/DDK-tagged full-length human PIP4K2C (hPIP4K2C-Myc/DDK), using PIP4K2C (E7P3S) Rabbit mAb (upper), HA-Tag (C29F4) Rabbit mAb #3724 (upper-middle), Myc-Tag (71D10) Rabbit mAb #2278 (lower-middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunoprecipitation Image 1: PIP4K2C (E7P3S) Rabbit mAb Expand Image
Immunoprecipitation of PIP4K2C protein from SHP-77 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is PIP4K2C (E7P3S) Rabbit mAb. Western blot analysis was performed using PIP4K2C (E7P3S) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
Immunohistochemistry Image 1: PIP4K2C (E7P3S) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human gastric adenocarcinoma using PIP4K2C (E7P3S) Rabbit mAb.
Immunohistochemistry Image 2: PIP4K2C (E7P3S) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human papillary thyroid carcinoma using PIP4K2C (E7P3S) Rabbit mAb.
Immunohistochemistry Image 3: PIP4K2C (E7P3S) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using PIP4K2C (E7P3S) Rabbit mAb.
Immunohistochemistry Image 4: PIP4K2C (E7P3S) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal human kidney using PIP4K2C (E7P3S) Rabbit mAb.
Immunohistochemistry Image 5: PIP4K2C (E7P3S) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal human testis using PIP4K2C (E7P3S) Rabbit mAb.
Immunohistochemistry Image 6: PIP4K2C (E7P3S) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal human stomach using PIP4K2C (E7P3S) Rabbit mAb.
Immunohistochemistry Image 7: PIP4K2C (E7P3S) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded Renca syngeneic tumor using PIP4K2C (E7P3S) Rabbit mAb.
Immunohistochemistry Image 8: PIP4K2C (E7P3S) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse brain using PIP4K2C (E7P3S) Rabbit mAb.
Immunohistochemistry Image 9: PIP4K2C (E7P3S) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse testis using PIP4K2C (E7P3S) Rabbit mAb.
Immunohistochemistry Image 10: PIP4K2C (E7P3S) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using PIP4K2C (E7P3S) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right). 
Immunohistochemistry Image 11: PIP4K2C (E7P3S) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded COLO 201 cell pellet (left, positive) or REH cell pellet (right, negative) using PIP4K2C (E7P3S) Rabbit mAb.
Immunofluorescence Image 1: PIP4K2C (E7P3S) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of fixed frozen mouse cerebellum labeled with PIP4K2C (E7P3S) Rabbit mAb (left, green). Free secondary binding sites were then blocked with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 prior to co-labeling with AQP4 (D1F8E) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #89851 (red), GFAP (GA5) Mouse mAb (Alexa Fluor® 555 Conjugate) #3656 (magenta), and DAPI #4083 (right, blue).
Immunofluorescence Image 1: PIP4K2C (E7P3S) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of Colo 201 cells (left, positive) or REH cells (right, negative) using PIP4K2C (E7P3S) Rabbit mAb (green) and DAPI #4083 (blue).
Flow Cytometry Image 1: PIP4K2C (E7P3S) Rabbit mAb Expand Image
Flow cytometric analysis of TALL-1 cells (blue, negative) and REC-1 cells (green, positive) using PIP4K2C (E7P3S) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.