Revision 3

#17336

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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.

Product Includes Product # Quantity Mol. Wt Isotype/Source
AHCYL1/IRBIT (D3A5G) Rabbit Monoclonal Antibody9424820 µl61 kDaRabbit IgG
CBS (D8F2P) Rabbit Monoclonal Antibody1478220 µl61 kDaRabbit IgG
Cystathionine gamma-Lyase (D1N1D) Rabbit Monoclonal Antibody1968920 µl44 kDaRabbit IgG
MTHFR (D1E4V) Rabbit Monoclonal Antibody2516420 µl78 kDaRabbit IgG
MTHFD1L (D8T7L) Rabbit Monoclonal Antibody1499920 µl106 kDaRabbit IgG
MTHFD2 (D8W9U) Rabbit Monoclonal Antibody4137720 µl35 kDaRabbit IgG
SHMT1 (D3B3J) Rabbit mAb8071520 µl50 kDaRabbit IgG
Thymidylate Synthase (D5B3) Rabbit Monoclonal Antibody904520 µl30 kDaRabbit
Anti-rabbit IgG, HRP-linked Antibody7074100 µlGoat

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

Description

The One-Carbon Metabolism Antibody Sampler Kit provides an economical means of detecting select components involved in one-carbon metabolism pathway. 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

One-carbon metabolism includes various enzymatic reactions involving the transfer of one-carbon groups mediated by folate cofactor (1). The activated one-carbon groups are used by various metabolic pathways, including purine synthesis, thymidine synthesis, and remethylation of homocysteine to methionine (1). S-adenosylhomocysteine hydrolase-like protein 1 (AHCYL1) is a member of the S-adenosylhomocysteine hydrolase family, which participates in the metabolism of S-adenosyl-L-homocysteine (2). Cystathionine beta-synthase (CBS) is a key enzyme involved in sulfur amino acid metabolism as it catalyzes the formation of cystathionine from serine and homocysteine (3,4). Cystathionine γ-lyase (CGL) is an enzyme in the transsulfuration pathway, a route in the metabolism of sulfur-containing amino acids (5). Methylenetetrahydrofolate reductase (MTHFR), a key enzyme in one-carbon metabolism, catalyzes the conversion of 5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate (1). 5-methyltetrahydrofolate donates its methyl group for remethylation of homocysteine to methionine (1). Methionine is further converted to S-adenosylmethionine (SAM), a major reactive methyl carrier (1). NADP+ dependent methylenetetrahydrofolate dehydrogenase 1-like (MTHFD1L) is a mitochondrial enzyme that catalyzes the production of formate from 10-formyl-tetrahydrofolate in one-carbon flow from mitochondria to cytoplasm (6,7). MTHFD2 is a bifunctional methylenetetrahydrofolate dehydrogenase/cyclohydrolase involved in mitochondrial folate metabolism (8). Serine hydroxymethyltransferase 1 (SHMT1) is a cytoplasmic serine hydroxylmethyltransferase (9,10). It catalyzes the conversion of serine to glycine with the transfer of β-carbon from serine to tetrahydrofolate (THF) to form 5, 10-methylene-THF (9, 10). The methylation of deoxyuridine monophosphate (dUMP) to deoxythymidine monophosphate (dTMP) is an essential step in the formation of thymine nucleotides, a process catalyzed by thymidylate synthase (TS or TYMS) (11-13).

Trademarks and Patents

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

CST is a registered 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 3

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using CBS (D8F2P) Rabbit mAb.
undefined Image 1: One-Carbon Metabolism Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using MTHFD1L (D8T7L) Rabbit mAb.
undefined Image 2: One-Carbon Metabolism Antibody Sampler Kit
Western blot analysis of extracts from K-562 cells, mouse liver, and rat liver using Cystathionine γ-Lyase (D1N1D) Rabbit mAb.
undefined Image 3: One-Carbon Metabolism 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 3

Cell Signaling Technology Logo
Western blot analysis of extracts from Daudi, Raji, and MOLT-4 cells using MTHFR (D1E4V) Rabbit mAb.
undefined Image 4: One-Carbon Metabolism Antibody Sampler Kit
Western blot analysis of extracts from HeLa and 293 cells using MTHFD2 (D8W9U) Rabbit mAb.
undefined Image 5: One-Carbon Metabolism 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 6: One-Carbon Metabolism 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 3

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using SHMT1 (D3B3J) Rabbit mAb.
undefined Image 7: One-Carbon Metabolism Antibody Sampler Kit
Western blot analysis of extracts from SK-BR-3 and HCC827 cells, untreated or treated with 5-FU (1 μM) or lapatinib (1 μM) for 24 hr, using Thymidylate Synthase (D5B3) XP® Rabbit mAb (upper) or β-Actin (13E5) Rabbit mAb #4970 (lower).
undefined Image 8: One-Carbon Metabolism Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using AHCYL1/IRBIT (D3A5G) Rabbit mAb.
undefined Image 9: One-Carbon Metabolism 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 3

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using Thymidylate Synthase (D5B3) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb (lower) #8457.
undefined Image 10: One-Carbon Metabolism Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Thymidylate Synthase (D5B3) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
undefined Image 11: One-Carbon Metabolism Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human lymphoma using Thymidylate Synthase (D5B3) XP® Rabbit mAb.
undefined Image 12: One-Carbon Metabolism 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 3

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded mouse spleen using Thymidylate Synthase (D5B3) XP® Rabbit mAb.
undefined Image 13: One-Carbon Metabolism Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded SK-BR-3 cell pellets, untreated (left) or treated with LY294002 #9901 (right), using Thymidylate Synthase (D5B3) XP® Rabbit mAb.
undefined Image 14: One-Carbon Metabolism Antibody Sampler Kit
Confocal immunofluorescent analysis of LN18 cells using Thymidylate Synthase (D5B3) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
undefined Image 15: One-Carbon Metabolism 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 3

Cell Signaling Technology Logo
Flow cytometric analysis of HCC827 cells (blue) and HeLa cells (green) using Thymidylate Synthase (D5B3) XP® Rabbit mAb.
undefined Image 16: One-Carbon Metabolism 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.