Biochemistry lab reports writing

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I_ hL· -..11wlc cxcl11sive_ c11;1y111~ o f the: cycle i'> s uccinatc dchyd,u~ena"c (Sl)f-IJ th<t l l'.-- t1gl1tl) hound to the 11111cr 1111tochonclr1al mcmbrnnc.

Steps of the TCA Cycle

{ IJL· I ( ' A cycle takes place: over eight different sleps:

• Step 1: f-ir~t the acctyl CoA (a two carbon molecule) joins with oxaloacetate (4 carbon inolccultJ to forn1 citrate (6 carbon molecule) Step 2: n,e citrate j::, then converted t(l i ·; oc i trr1.tc (i~rnner of citn,.u-) Step J: fsocitrnl.e; 1~ then oxicli:-:.ed to alpha-kctoglutarate (a five carbon molecule) which : ec.,u I 1 ') i 1 tli-: rck,1>0..: of c1rbon dioxide One NADH molecule is also formed in this step.

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Dct ~rmi 11 ,ltio11 o f ac ti , it) of k rc h s C) c lc dch:-, d rogc n as c s in th e li ver.

f) '-'h: dr1..1gcn:1~1..· nct 1\ it:-, 1::- 1..· a rricd ou t by a tetra z,) l1um me thod:

Dehydrogcnases arc the en 7: mes o xo id i7 ing the appro priate s u bs tra tes . 2.3 .5-L iph~n:-, l- telr.1 L0liu111 chloride is uc:;e<l as a hyd ro ge n acc e ;Ho r w hi c h is red u ced a nd fo rm s the~ co lored gn' wet. [ntens ity of the co lo r de pe nd s on th e am o u n t o f the fo r m ed product c:1u d -.:o n seq uc nt l: on deh yd ro genasc activ ity P:-, ru , ·ate . · ._ o cJtrntc.2-o , o glLtteratc,succinate.and m a la tc are u sed as the substrate $_.

Cr eneral chemical ~,1uat ;nn is as shov.•n bel o\,

Sub,·1ra1e +tetrazo/i u m (crJ/o rless) ____ _

Proce d ure

__de t etraz.oli um(re d c olored)

hy drog e nated p ro duc t

+redu ced su bstr a te

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Determiu ation of succinate dehydrogcnasc acti, ity in , arious tissues/

r nhihition of s uc c inate dchyd rogenase by azidc/q anide

~ucci11ate clchydrogcn,1 sc reactio n is il lu strated by the fo ll O\ving. equation.

Succ inate+ f>ID or tetrnzolium 1mwrate +FADl/2 or ----------'

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Reduced tetrazo/ium

I. liomogcnates of the brain, heart. kidney s. liver, skeletal muscle. and \unf are used to work as a source of succinate dehyc\rogenase.

2. Prepare the incubation mi xt ures as follows 3. Mix th e contents and incubate at 3 8 degrees for 45 rnins

-- - -- - T -- Estirnat•~

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