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17336
One-Carbon Metabolism Antibody Sampler Kit

One-Carbon Metabolism Antibody Sampler Kit #17336

Western Blotting Image 1

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

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Western Blotting Image 2

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

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Western Blotting Image 3

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

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Western Blotting Image 4

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

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Western Blotting Image 5

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

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Western Blotting Image 6

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

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Western Blotting Image 7

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

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Western Blotting Image 8

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).

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Western Blotting 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.

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Western Blotting Image 10

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.

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IHC-P (paraffin) 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).

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IHC-P (paraffin) Image 12

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

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IHC-P (paraffin) Image 13

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

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IHC-P (paraffin) 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.

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Flow Cytometry Image 15

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

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IF-IC Image 16

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

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
AHCYL1/IRBIT (D3A5G) Rabbit mAb 94248 20 µl
  • WB
  • IP
H M R Mk 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 M R Mk 50 Rabbit IgG
Thymidylate Synthase (D5B3) XP® Rabbit mAb 9045 20 µl
  • WB
  • IHC
  • IF
  • F
H M R Mk 30 Rabbit 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

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.

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.

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.

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, 43178-43187.
  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.
Entrez-Gene Id
10768 , 875 , 1491 , 25902 , 10797 , 4524 , 6470 , 7298
Swiss-Prot Acc.
O43865 , P35520 , P32929 , Q6UB35 , P13995 , P42898 , P34896 , P04818
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.

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