Anti-ACADM/MCAD antibody [3B7BH7] (ab110296)

Vielen Dank für Ihre Antwort.

Leider ist mir bei meiner letzten Antwort ein Fehler unterlaufen und ich möchte diesen hiermit korrigieren:

ab110296 wird nicht im Kit ACADM ELISA Kit (ab129734) verwendet.. Es wird ein Antikörper verwendet, der nicht einzeln erhältlich ist. Bitte entschuldigen Sie vielmals diese Fehlinformation.

Bezüglich der anderen Fragen habe ich unsere Spezialisten bei MitoSciences gefragt. Ich habe ihnen die Antwort direkt unten rein kopiert:

"We purchased the cell line with the K304E (984A>G in ACADM gene) from the Coriell repository. The cell line is homozygous for 984A>G . The mutation in this cell line was found with a decrease in MCAD activity (12% from control) by the submitter, but there is no specific reference for this particular cell line in the coriell website.

http://ccr.coriell.org/Sections/Search/Sample_Detail.aspx?Ref=GM07844&PgId=166



However, it is well known in the literature that the K304E aa change (homozygous or compound heterozygous) induces MCAD deficiency (measured by enzyme activity). References below. Many of these studies were done before there were commercially available MCAD antibodies. So the WB data does not exist. Therefore we tested the above cell line from Coriell with our antibody 3B7BH7 and found that MCAD protein levels are significantly decreased (both by WB and by ICC). The customer can look at the data in the ICE booklet as mentioned in my previous email. We don’t have WB data with the antibody used in the kit showing control vs deficient, because the antibody is WB negative.

A good place to look at mutations known to induce MCAD deficiency is OMIM.

http://omim.org/entry/201450



http://omim.org/entry/201450#reference32 reported the findings in a prospective neonatal screening program in Pennsylvania using tandem mass spectrometry. From the first 80,371 newborns screened, they found 9 babies with MCAD (1/8930), plus 2 additional newborns, screened because of a previously known family history. Molecular analysis showed that 56% of the patients were compound heterozygotes for the 985A-G mutation (K304E; http://omim.org/entry/607008#0001), commonly referred to as G985, and a second mutation.


In 9 patients with MCAD deficiency, http://omim.org/entry/201450#reference16 found a 985A-G transition which resulted in a lys304-to-glu substitution (K304E; http://omim.org/entry/607008#0001) in the mature protein. These patients were unrelated, suggesting a high incidence of this abnormality among Caucasian patients. The change was not found in 20 healthy Caucasian and 6 healthy Japanese subjects. http://omim.org/entry/201450#reference16 found this point mutation in 31 of 34 (91%) mutant MCAD alleles. http://omim.org/entry/201450#reference31 estimated that 90% of MCAD cases involve the same mutation.

http://omim.org/entry/201450#reference33 characterized the molecular defect in 4 patients with mild MCAD deficiency. In routine neonatal screening on the fifth day of life, they had been found to have abnormal acylcarnitine profiles indicative of MCAD deficiency. Two were of German origin and the other 2 were born to different consanguineous Turkish parents. In all 4, the clinical course and routine laboratory investigations up to the age of 6 months were unremarkable. Enzyme studies showed residual MCAD activities between those with classic MCAD deficiency and heterozygotes. In 2 cases, ACADM gene analysis revealed compound heterozygosity for the common K304E mutation (http://omim.org/entry/607008#0001) and the 199T-C mutation (Y42H; http://omim.org/entry/607008#0011), which they designated Y67H. In the 2 children of consanguineous parents, homozygosity was found for the gly267-to-arg mutation (G267R; http://omim.org/entry/607008#0003) and the S220L mutation (http://omim.org/entry/607008#0012), respectively. As in other metabolic disorders, the distinction between 'normal' and 'disease' in MCAD deficiency is blurred into a spectrum of enzyme deficiency states caused by different mutations in the ACADM gene potentially influenced by factors affecting intracellular protein processing."



Ich hoffe, diese Information ist hilfreich und wird Ihnen Ihre Bedenken bezüglich des Kits nehmen.

Vielen Dank für Ihre Anfrage.

Das Immunogen für ab110296 war das Protein voller Länge. Das Epitop, das der Antikörper bindet ist leider nicht näher kartiert worden und wir kennen es deshalb nicht.

Ich freue mich Ihnen bestätigen zu können, dass ab110296 im Kit ACADM ELISA Kit (ab129734) verwendet wird.

Es tut mir leid, dass ich Ihre Fragen diesmal nicht vollständig beantworten konnte und hoffe, dass diese Information dennoch hilfreich ist.