MMACHC Monoclonal antibody

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  WB analysis of HepG2 using 66609-1-Ig

  HepG2 cells were subjected to SDS PAGE followed by western blot with 66609-1-Ig (MMACHC antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis of Raji using 66609-1-Ig

  Raji cells were subjected to SDS PAGE followed by western blot with 66609-1-Ig (MMACHC antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis of K-562 using 66609-1-Ig

  K-562 cells were subjected to SDS PAGE followed by western blot with 66609-1-Ig (MMACHC antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis of RAW 264.7 using 66609-1-Ig

  RAW 264.7 cells were subjected to SDS PAGE followed by western blot with 66609-1-Ig (MMACHC antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis of HEK-293 using 66609-1-Ig

  HEK-293 cells were subjected to SDS PAGE followed by western blot with 66609-1-Ig (MMACHC antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HeLa using 66609-1-Ig

  HeLa cells were subjected to SDS PAGE followed by western blot with 66609-1-Ig (MMACHC antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  IF Staining of HepG2 using 66609-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using MMACHC antibody (66609-1-Ig, Clone: 4G10A11 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L), CL594-Phalloidin (red).

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  FC experiment of HeLa using 66609-1-Ig

  1X10^6 HeLa cells were intracellularly stained with 0.8 ug Anti-Human MMACHC (66609-1-Ig, Clone:4G10A11) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.8 ug Mouse IgG1 Isotype Control (MOPC-21) (65124-1-Ig, Clone: MOPC-21) (blue). Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

MMACHC Monoclonal Antibody for WB, IF/ICC, FC (Intra), ELISA

Cat No. 66609-1-Ig

Clone No.4G10A11

Publications(2)

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse and More (1)

Applications

WB, IF/ICC, FC (Intra), ELISA

Synonyms

4G10A11, Alkylcobalamin:glutathione S-alkyltransferase, CblC, cyanide-eliminating, Cyanocobalamin reductase

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Tested Applications

• Positive WB detected in: HepG2 cells, K-562 cells, HEK-293 cells, HeLa cells, Raji cells, RAW 264.7 cells
• Positive IF/ICC detected in: HepG2 cells
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.80 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 2 publications below

Product Information

66609-1-Ig targets MMACHC in WB, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human, mouse samples.

• Tested Reactivity: human, mouse
• Cited Reactivity: human, rat
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen:

  CatNo: Ag15001

  Product name: Recombinant human MMACHC protein

  Source: e coli.-derived, PET28a

  Tag: 6*His

  Domain: 1-225 aa of BC006122

  Sequence: MFDRALKPFLQSCHLRMLTDPVDQCVAYHLGRVRESLPELQIEIIADYEVHPNRRPKILAQTAAHVAGAAYYYQRQDVEADPWGNQRISGVCIHPRFGGWFAIRGVVLLPGIEVPDLPPRKPHDCVPTRADRIALLEGFNFHWRDWTYRDAVTPQERYSEEQKAYFSTPPAQRLALLGLAQPSEKPSSPSPDLPFTTPAPKKPGNPSRARSWLSPRVSPPASPGP

• Full Name: methylmalonic aciduria (cobalamin deficiency) cblC type, with homocystinuria
• Calculated Molecular Weight: 282 aa, 32 kDa
• Observed Molecular Weight: 32 kDa
• GenBank Accession Number: BC006122
• Gene Symbol: MMACHC
• Gene ID (NCBI): 25974
• RRID: AB_2881969
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• UNIPROT ID: Q9Y4U1
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

MMACHC, also named as CblC, is located on chromosome 1 and comprises four coding exons. Human MMACHC catalyzes the reductive decyanation of cyanocobalamin and may serve as a trafficking chaperone for intracellular cobalamin. It also catalyzes the glutathione-dependent reductive demethylation of methylcobalamin, and, with much lower efficiency, the glutathione-dependent reductive demethylation of adenosylcobalamin. NMACHC binds cyanocobalamin, adenosylcobalamin, methylcobalamin and other, related vitamin B12 derivatives.

Publications

• Human
• Mouse
• Rat
• Monkey
• Pig

Species	Application	Title
human	WB	Cancer Lett; CYLD deficiency enhances metabolic reprogramming and tumor progression in nasopharyngeal carcinoma via PFKFB3.; Authors - Lingzhi Wang
rat	WB	Mol Neurobiol; Surfeit Locus Protein 4 as a Novel Target for Therapeutic Intervention in Cerebral Ischemia-Reperfusion Injury; Authors - Wenjie Hu