Revision 1

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877-616-CELL (2355)

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Support:

877-678-TECH (8324)

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

Product Includes Product # Quantity Mol. Wt Isotype/Source
Fumarase (D9C5) Rabbit Monoclonal Antibody456720 µl49 kDaRabbit IgG
IDH2 (D8E3B) Rabbit Monoclonal Antibody5643920 µl43 kDaRabbit IgG
Citrate Synthase (D7V8B) Rabbit Monoclonal Antibody1430920 µl45 kDaRabbit IgG
ACO2 (D6D9) Rabbit Monoclonal Antibody657120 µl85 kDaRabbit IgG
MPC2 (D4I7G) Rabbit Monoclonal Antibody4614120 µl14 kDaRabbit IgG
MPC1 (D2L9I) Rabbit Monoclonal Antibody1446220 µl12 kDaRabbit IgG
DLST (D22B1) Rabbit Monoclonal Antibody1195420 µl50 kDaRabbit IgG
SDHA (D6J9M) Rabbit Monoclonal Antibody1199820 µl70 kDaRabbit IgG
Anti-rabbit IgG, HRP-linked Antibody7074100 µlGoat
IDH1 Antibody399720 µl46 kDaRabbit

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Tricarboxylic Acid Cycle Sampler Kit provides an economical means of detecting select components involved in tricarboxylic acid cycle. The kit contains enough primary antibodies to perform at least two western blot experiments per antibody.

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.

Background

The tricarboxylic acid (TCA) cycle includes various enzymatic reactions that constitute a key part of cellular aerobic respiration. The transport of the glycolytic end product pyruvate into mitochondria and the decarboxylation of pyruvate in the TCA cycle generate energy through oxidative phosphorylation under aerobic conditions (1,2). Two inner mitochondrial membrane proteins, mitochondrial pyruvate carrier 1 (MPC1) and mitochondrial pyruvate carrier 2 (MPC2), form a 150 kDa complex and are essential proteins in the facilitated transport of pyruvate into mitochondria (1,2). Citrate synthase catalyzes the first and rate-limiting reaction of the TCA cycle (3). Mitochondrial aconitase 2 (ACO2) catalyzes the conversion of citrate to isocitrate via cis-aconitate (4). IDH1 and IDH2 are two of the three isocitrate dehydrogenases that catalyze oxidative decarboxylation of isocitrate to α-ketoglutarate (α-KG) (5). IDH1 functions as a tumor suppressor in the cytoplasm and peroxisomes, whereas IDH2 is in mitochondria and is involved in the TCA cycle (5). Mutations in IDH2 have also been identified in malignant gliomas (6). Dihydrolipoamide succinyltransferase (DLST) is a subunit of the α-ketoglutarate dehydrogenase complex, a key enzymatic complex in the TCA cycle (7). Succinate dehydrogenase subunit A (SDHA) is a component of the TCA cycle and the electron transport chain and is involved in the oxidation of succinate (8). Fumarase catalyzes the conversion of fumarate to malate (9). Fumarase deficiency leads to the accumulation of fumarate, an oncometabolite that has been shown to promote epithelial-to-mesenchymal-transition (EMT), a developmental process that has been implicated in oncogenesis (10).

Trademarks and Patents

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

SignalFire is a registered 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 1

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using DLST (D22B1) XP® Rabbit mAb.
undefined Image 1: Tricarboxylic Acid Cycle Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using SDHA (D6J9M) XP® Rabbit mAb.
undefined Image 2: Tricarboxylic Acid Cycle Antibody Sampler Kit
Flow cytometric analysis of HeLa cells using SDHA (D6J9M) XP® Rabbit mAb (solid line) compared to 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) was used as a secondary antibody.
undefined Image 3: Tricarboxylic Acid Cycle Antibody Sampler Kit
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 1

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using Citrate Synthase (D7V8B) Rabbit mAb.
undefined Image 4: Tricarboxylic Acid Cycle Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using MPC1 (D2L9I) Rabbit mAb.
undefined Image 5: Tricarboxylic Acid Cycle Antibody Sampler Kit
Western blot analysis of extracts from HepG2 and NIH/3T3 cells using IDH1 Antibody.
undefined Image 6: Tricarboxylic Acid Cycle Antibody Sampler Kit
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 1

Cell Signaling Technology Logo
Western blot analysis of extracts from 293 cells, untransfected (-) or transfected with a Myc-tagged cDNA expression construct encoding human fumarase (hFumarase-Myc; +), using Fumarase (D9C5) Rabbit mAb (upper) and Myc-Tag (71D10) Rabbit mAb #2278 (lower).
undefined Image 7: Tricarboxylic Acid Cycle Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using Fumarase (D9C5) Rabbit mAb.
undefined Image 8: Tricarboxylic Acid Cycle Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using MPC2 (D4I7G) Rabbit mAb.
undefined Image 9: Tricarboxylic Acid Cycle Antibody Sampler Kit
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 1

Cell Signaling Technology Logo
Western blot analysis of extracts from MCF7 and ACHN cells using IDH2 (D8E3B) Rabbit mAb.
undefined Image 10: Tricarboxylic Acid Cycle Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using ACO2 (D6D9) XP® Rabbit mAb.
undefined Image 11: Tricarboxylic Acid Cycle Antibody Sampler Kit
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
undefined Image 12: Tricarboxylic Acid Cycle Antibody Sampler Kit
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 1

Cell Signaling Technology Logo
Immunoprecipitation of DLST from HeLa cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or DLST (D22B1) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using DLST (D22B1) XP® Rabbit mAb. Mouse Anti-rabbit IgG (Light-Chain Specific) (L57A3) mAb #3677 and Anti-mouse IgG, HRP-linked Antibody #7076 were used as secondary antibodies.
undefined Image 13: Tricarboxylic Acid Cycle Antibody Sampler Kit
Immunoprecipitation of SDHA from HeLa cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or SDHA (D6J9M) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using SDHA (D6J9M) XP® Rabbit mAb.
undefined Image 14: Tricarboxylic Acid Cycle Antibody Sampler Kit
Flow cytometric analysis of C2C12 cells using SDHA (D6J9M) XP® Rabbit mAb (solid line) compared to 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) was used as a secondary antibody.
undefined Image 15: Tricarboxylic Acid Cycle Antibody Sampler Kit
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 1

Cell Signaling Technology Logo
Confocal immunofluorescent analysis of HeLa cells using Citrate Synthase (D7V8B) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
undefined Image 16: Tricarboxylic Acid Cycle Antibody Sampler Kit
Western blot analysis of extracts from 293 cells, mock transfected (-) or transfected with a construct expressing DDK-tagged full-length human MPC1 protein (hMPC1-DDK; +) or HA-tagged full-length human MPC2 protein (hMPC2-HA; +), using MPC1 (D2L9I) Rabbit mAb (upper), DYKDDDDK Tag (9A3) Mouse mAb #8146 (upper middle), HA-Tag (C29F4) Rabbit mAb #3724 (lower middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower). Both expression vectors were provided by Dr. Jared Rutter's laboratory at the University of Utah School of Medicine.
undefined Image 17: Tricarboxylic Acid Cycle Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using Fumarase (D9C5) Rabbit mAb.
undefined Image 18: Tricarboxylic Acid Cycle Antibody Sampler Kit
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 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human thymoma using Fumarase (D9C5) Rabbit mAb.
undefined Image 19: Tricarboxylic Acid Cycle Antibody Sampler Kit
Immunoprecipitation of IDH2 from Hep G2 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is IDH2 (D8E3B) Rabbit mAb. Western blot analysis was performed using IDH2 (D8E3B) Rabbit mAb.
undefined Image 20: Tricarboxylic Acid Cycle Antibody Sampler Kit
Confocal immunofluorescent analysis of HeLa cells using ACO2 (D6D9) XP® Rabbit mAb (green), showing colocalization with mitochondria that were labeled with MitoTracker® Red CMXRos (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
undefined Image 21: Tricarboxylic Acid Cycle Antibody Sampler Kit
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 1

Cell Signaling Technology Logo
Confocal immunofluorescent analysis of HeLa cells using DLST (D22B1) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
undefined Image 22: Tricarboxylic Acid Cycle Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using SDHA (D6J9M) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
undefined Image 23: Tricarboxylic Acid Cycle Antibody Sampler Kit
Immunoprecipitation of MPC1 from Huh7 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or MPC1 (D2L9I) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using MPC1 (D2L9I) Rabbit mAb.
undefined Image 24: Tricarboxylic Acid Cycle Antibody Sampler Kit
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 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded normal human liver using Fumarase (D9C5) Rabbit mAb.
undefined Image 25: Tricarboxylic Acid Cycle Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using IDH2 (D8E3B) Rabbit mAb.
undefined Image 26: Tricarboxylic Acid Cycle Antibody Sampler Kit
Confocal immunofluorescent analysis of HeLa cells using SDHA (D6J9M) XP® Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
undefined Image 27: Tricarboxylic Acid Cycle Antibody Sampler Kit
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 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded normal human kidney using Fumarase (D9C5) Rabbit mAb.
undefined Image 28: Tricarboxylic Acid Cycle Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded K-562 cell pellet (left, high-expressing) or SR cell pellet (right, low-expressing) using IDH2 (D8E3B) Rabbit mAb.
undefined Image 29: Tricarboxylic Acid Cycle Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded normal human adrenal gland using Fumarase (D9C5) Rabbit mAb.
undefined Image 30: Tricarboxylic Acid Cycle Antibody Sampler Kit
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 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using IDH2 (D8E3B) Rabbit mAb.
undefined Image 31: Tricarboxylic Acid Cycle Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded 4T1 syngeneic tumor using Fumarase (D9C5) Rabbit mAb.
undefined Image 32: Tricarboxylic Acid Cycle Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human adenoid cystic carcinoma of the trachea using IDH2 (D8E3B) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
undefined Image 33: Tricarboxylic Acid Cycle Antibody Sampler Kit
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 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded mouse colon using Fumarase (D9C5) Rabbit mAb.
undefined Image 34: Tricarboxylic Acid Cycle Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma using IDH2 (D8E3B) Rabbit mAb.
undefined Image 35: Tricarboxylic Acid Cycle Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded mouse spleen using Fumarase (D9C5) Rabbit mAb.
undefined Image 36: Tricarboxylic Acid Cycle Antibody Sampler Kit
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 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using IDH2 (D8E3B) Rabbit mAb.
undefined Image 37: Tricarboxylic Acid Cycle Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human small intestine using Fumarase (D9C5) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
undefined Image 38: Tricarboxylic Acid Cycle Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using IDH2 (D8E3B) Rabbit mAb.
undefined Image 39: Tricarboxylic Acid Cycle Antibody Sampler Kit
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 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded 293T cell pellet, untransfected (left) or fumarase-transfected (right), using Fumarase (D9C5) Rabbit mAb.
undefined Image 40: Tricarboxylic Acid Cycle Antibody Sampler Kit
Flow cytometric analysis of NIH/3T3 cells using Citrate Synthase (D7V8B) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
undefined Image 41: Tricarboxylic Acid Cycle Antibody Sampler Kit
Flow cytometric analysis of KMS-11 cells using Citrate Synthase (D7V8B) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
undefined Image 42: Tricarboxylic Acid Cycle Antibody Sampler Kit
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.