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17336
One-Carbon Metabolism Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

One-Carbon Metabolism Antibody Sampler Kit #17336

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One-Carbon Metabolism Antibody Sampler Kit: Image 1

Western blot analysis of extracts from various cell lines using AHCYL1/IRBIT (D3A5G) Rabbit mAb.

One-Carbon Metabolism Antibody Sampler Kit: Image 2

Western blot analysis of extracts from various cell lines using CBS (D8F2P) Rabbit mAb.

One-Carbon Metabolism Antibody Sampler Kit: Image 3

Western blot analysis of extracts from K-562 cells, mouse liver, and rat liver using Cystathionine γ-Lyase (D1N1D) Rabbit mAb.

One-Carbon Metabolism Antibody Sampler Kit: Image 4

Western blot analysis of extracts from Daudi, Raji, and MOLT-4 cells using MTHFR (D1E4V) Rabbit mAb.

One-Carbon Metabolism Antibody Sampler Kit: Image 5

Western blot analysis of extracts from various cell lines using MTHFD1L (D8T7L) Rabbit mAb.

One-Carbon Metabolism Antibody Sampler Kit: Image 6

Western blot analysis of extracts from HeLa and 293 cells using MTHFD2 (D8W9U) Rabbit mAb.

One-Carbon Metabolism Antibody Sampler Kit: Image 7

Western blot analysis of extracts from various cell lines using SHMT1 (D3B3J) Rabbit mAb.

One-Carbon Metabolism Antibody Sampler Kit: Image 8

Flow cytometric analysis of HCC827 cells (blue) and HeLa cells (green) using Thymidylate Synthase (D5B3) XP® Rabbit mAb.

One-Carbon Metabolism Antibody Sampler Kit: Image 9

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.

One-Carbon Metabolism Antibody Sampler Kit: Image 10

Confocal immunofluorescent analysis of LN18 cells using Thymidylate Synthase (D5B3) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

One-Carbon Metabolism Antibody Sampler Kit: Image 11

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).

One-Carbon Metabolism Antibody Sampler Kit: Image 12

Immunohistochemical analysis of paraffin-embedded human lymphoma using Thymidylate Synthase (D5B3) XP® Rabbit mAb.

One-Carbon Metabolism Antibody Sampler Kit: Image 13

Immunohistochemical analysis of paraffin-embedded mouse spleen using Thymidylate Synthase (D5B3) XP® Rabbit mAb.

One-Carbon Metabolism Antibody Sampler Kit: Image 14

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.

One-Carbon Metabolism Antibody Sampler Kit: Image 15

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).

One-Carbon Metabolism Antibody Sampler Kit: Image 16

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.

To Purchase # 17336T
Product # Size Price
17336T
1 Kit  (8 x 20 µl) $ 554

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
AHCYL1/IRBIT (D3A5G) Rabbit mAb 94248 20 µl
  • WB
  • IP
H Mk M R 61 Rabbit IgG
CBS (D8F2P) Rabbit mAb 14782 20 µl
  • WB
H M R 61 Rabbit IgG
Cystathionine γ-Lyase (D1N1D) Rabbit mAb 19689 20 µl
  • WB
H M R 44 Rabbit IgG
MTHFR (D1E4V) Rabbit mAb 25164 20 µl
  • WB
H 78 Rabbit IgG
 MTHFD1L (D8T7L) Rabbit mAb 14999 20 µl
  • WB
H M 106 Rabbit IgG
MTHFD2 (D8W9U) Rabbit mAb 41377 20 µl
  • WB
  • IP
H 35 Rabbit IgG
SHMT1 (D3B3J) Rabbit mAb 80715 20 µl
  • WB
  • IP
H Mk M R 50 Rabbit IgG
Thymidylate Synthase (D5B3) XP® Rabbit mAb 9045 20 µl
  • WB
  • IHC
  • IF
  • F
H Mk M R 30 Rabbit 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product 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.

Specificity / Sensitivity

Each antibody in this kit recognizes endogenous levels of its specific target protein. AHCYL1/IRBIT (D3A5G) Rabbit mAb does not cross-react with AHCYL2/IRBIT2 protein. SHMT1 (D3B3J) rabbit mAb does not cross-react with SHMT2 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Glu50 of human AHCYL1/IRBIT protein, Leu220 of human CBS protein, Pro298 of human cystathionine γ-lyase protein, Pro39 of human MTHFR protein, Ala264 of human MTHFD1L protein, Leu340 of human MTHFD2 protein, Glu187 of human SHMT1 protein, and residues near the carboxy terminus of human thymidylate synthase protein.

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).

  1. Ducker, G.S. and Rabinowitz, J.D. (2017) Cell Metab 25, 27-42.
  2. Jeong, W. et al. (2012) PLoS One 7, e49204.
  3. Banerjee, R. and Zou, C.G. (2005) Arch Biochem Biophys 433, 144-56.
  4. Jhee, K.H. and Kruger, W.D. Antioxid Redox Signal 7, 813-22.
  5. Chiku, T. et al. (2009) J Biol Chem 284, 11601-12.
  6. Prasannan, P. et al. (2003) J Biol Chem 278, 43178-87.
  7. Prasannan, P. and Appling, D.R. (2009) Arch Biochem Biophys 481, 86-93.
  8. Christensen, K.E. and Mackenzie, R.E. (2008) Vitam Horm 79, 393-410.
  9. MacFarlane, A.J. et al. (2008) J Biol Chem 283, 25846-53.
  10. Hebbring, S.J. et al. (2012) J Neurochem 120, 881-90.
  11. Johnston, P.G. et al. (1991) Cancer Res 51, 6668-76.
  12. Aschele, C. et al. (2002) Ann Oncol 13, 1882-92.
  13. Jackman, A.L. and Calvert, A.H. (1995) Ann Oncol 6, 871-81.

Pathways & Proteins

Explore pathways + proteins related to this product.

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's 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's 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.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
CST is a trademark of Cell Signaling Technology, Inc.
SignalFire is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
LumiGLO is a registered trademark of Kirkegaard & Perry Laboratories.
Tween is a registered trademark of ICI Americas, Inc.