Revision 3

#97366

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, W-S, IP, IHC-P, FC-FP

Reactivity:
H M R

Sensitivity:
Endogenous

MW (kDa):
125

Source/Isotype:
Rabbit IgG

UniProt ID:
#P53396

Entrez-Gene Id:
47

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:50 - 1:250
Immunoprecipitation 1:100
Immunohistochemistry (Paraffin) 1:400 - 1:1600
Flow Cytometry (Fixed/Permeabilized) 1:400 - 1:1600

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

Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit Monoclonal Antibody recognizes endogenous levels of ATP-citrate lyase protein only when phosphorylated at Ser455 (equivalent to Ser454 in rat). Nonspecific plasma membrane staining was observed in a subset of tissues by immunohistochemistry.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser455 of human ATP-citrate lyase protein.

Background

ATP-citrate lyase (ACL) is a homotetramer that catalyzes the formation of acetyl-CoA and oxaloacetate (OAA) in the cytosol, which is the key step for the biosynthesis of fatty acids, cholesterol, and acetylcholine, as well as for gluconeogenesis (1). Nutrients and hormones regulate the expression level and phosphorylation of ATP-citrate lyase (1,2). It is phosphorylated by GSK-3 on Thr446 and Ser450 (3). Ser455 of ATP-citrate lyase has been reported to be phosphorylated by PKA and Akt (4,5). Phosphorylation on Ser455 abolishes the homotropic allosteric regulation by citrate and enhances the catalytic activity of the enzyme (2).

ATP-citrate lyase is also critical in the pathogenesis of Parkinson’s disease (PD). α-synuclein mutations activate ATP-citrate lyase, leading to elevated cytoplasmic p300 activity. Hyperactivation of cytoplasmic p300 subsequently promotes mTORC1 signaling to inhibit autophagy, causing the aggregation of α-synuclein (6).

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 nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

W: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: Human M: Mouse R: Rat

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
Western blot analysis of extracts from Jurkat cells, untreated (-) or treated with λ phosphatase and calf intestinal phosphatase (CIP) (+), using Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit mAb (upper), ATP-Citrate Lyase (D1X6P) Rabbit mAb #13390 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western Blotting Image 1: Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit Monoclonal Antibody
Western blot analysis of extracts from MCF7 cells, serum-starved and untreated (-) or treated with human IGF-I (hIGF-I) (100 ng/mL, 20 min; +) that were subsequently untreated (-) or treated with λ phosphatase and calf intestinal phosphatase (CIP) (+), using Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit mAb (upper), ATP-Citrate Lyase (D1X6P) Rabbit mAb #13390 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western Blotting Image 2: Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit Monoclonal Antibody
Western blot analysis of extracts from Jurkat cells, untreated (-) or treated with LY294002 #9901, Wortmannin #9951, and U0126 #9903 (50 μM, 1 μM, and 10 μM, 3 hr), using Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit mAb (upper), ATP-Citrate Lyase (D1X6P) Rabbit mAb #13390 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western Blotting Image 3: Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
Western blot analysis of extracts from Neuro-2a, EL4, and F9 cells using Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit mAb (upper), ATP-Citrate Lyase (D1X6P) Rabbit mAb #13390 (middle) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western Blotting Image 4: Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit Monoclonal Antibody
Western blot analysis of extracts from RBL-2H3, MTLn3, and C6 cells using Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit mAb (upper), ATP-Citrate Lyase (D1X6P) Rabbit mAb #13390 (middle) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western Blotting Image 5: Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit Monoclonal Antibody
Simple WesternTM analysis of lysates (0.1 mg/mL) from Neuro-2a cells usingPhospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit mAb #97366. The virtual lane view (left) shows the target (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the JessTM Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Western Blotting Image 1: Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
Immunoprecipitation of Phospho-ATP-Citrate Lyase protein from HeLa extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit mAb. Western blot analysis was performed using Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
Immunoprecipitation Image 1: Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit mAb.
Immunohistochemistry Image 1: Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded human esophageal adenocarcinoma using Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit mAb.
Immunohistochemistry Image 2: Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit mAb.
Immunohistochemistry Image 3: Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded human gastric adenocarcinoma using Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit mAb.
Immunohistochemistry Image 4: Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded normal human small intestine using Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit mAb.
Immunohistochemistry Image 5: Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded normal human kidney using Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit mAb.
Immunohistochemistry Image 6: Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded CT26.WT syngeneic tumor using Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit mAb.
Immunohistochemistry Image 7: Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded 4T1 syngeneic tumor using Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit mAb.
Immunohistochemistry Image 8: Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded mouse brain cortex using Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit mAb.
Immunohistochemistry Image 9: Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded mouse spleen using Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit mAb.
Immunohistochemistry Image 10: Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma, untreated (left) or lambda phosphatase treated (right), using Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit mAb.
Immunohistochemistry Image 11: Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma (left), serous papillary carcinoma of the ovary (center), or esophageal adenocarcinoma (right) using using Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit mAb (top) or ATP-Citrate Lyase (F4S7B) Rabbit mAb #73415 (bottom).
Immunohistochemistry Image 12: Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Immunohistochemistry Image 13: Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit Monoclonal Antibody
Flow cytometric analysis of Jurkat cells treated post-fixation with λ phosphatase (blue) or untreated (green), using Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit mAb (solid lines) or a 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.
Flow Cytometry Image 1: Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
Flow cytometric analysis of Jurkat cells treated with LY294002 #9901, Wortmannin #9951, and U0126 #9903 (50 μM, 1 μM, and 10 μM, 3 hr; blue, low-expressing) and untreated Jurkat cells (green, high-expressing) using Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit mAb (solid lines) or a 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.
Flow Cytometry Image 2: Phospho-ATP-Citrate Lyase (Ser455) (F4T8R) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.