RFE Fish Species Identification Isoelectric Focusing Tools

U.S. Food & Drug Administration
Seafood Products Research Center
Center for Food Safety & Applied Nutrition
Regulatory Fish Encyclopedia:
IEF Gel Lane Banding Pattern Peak Report Data

Peak Report Data from Isoelectric Focusing for a specimen of:
Orange Roughy ---- Go to Display Gel Lane Navigator.

FDA Seafood List Scientific Name: Hoplostethus atlanticus
FDA Seafood List Common Name: Orange Roughy
FDA Seafood List Market Name: Roughy, Orange
Unique IEF Gel Code: 19950802gel02
Unique Web Code: orh01Srr
Tissue Aliquot Code: hoplatla I94-001 #11a Gel Analysis Conditions


Gel Image	Fish Image	Calibrated Gel

DISPLAY GEL LANE NAVIGATOR
RETURN TO TOP OF THIS DOCUMENT
Lane a pI Markers	Lane b Specimen	Lane c Co-Migrated	Lane d pI Markers	Lane e Specimen	Lane f pI Markers	Go to Gel Schematic

**DISPLAY GEL LANE LETTER:** Lane a
**DIGITAL REPORT GEL LANE:** 2
**LANE TYPE:** pI Markers
**ACTUAL GEL LANE:** 2
Go to Gel Schematic
Return to Lane Navigator
Band	pI	Loc	% RF	Amplitude	Width	Integr. OD	% Integr. OD	% Lane IntOD	pI	G. Loc	G. Sigma	G. Ampl
1	4.55	89	10.2	0.0866409	6	0.919354	9.6	7.6	4.55	89.6	6.1	0.0852735
2	5.2	188	21.6	0.0821168	6	0.866694	9.1	7.1	5.2	187.3	6.1	0.0805716
3	5.85	303	34.9	0.0824875	5	0.696409	7.3	5.7	5.85	301.6	4.9	0.0802758
4	6.55	422	48.6	0.146361	6	1.42467	14.9	11.7	6.55	421.4	5.6	0.143256
5	6.85	457	52.6	0.0462878	5	0.408054	4.3	3.4	6.85	458.2	5.3	0.0435537
6	7.35	520	59.8	0.0849286	5	0.718382	7.5	5.9	7.35	519.3	4.9	0.0820659
7	8.15	646	74.3	0.119859	7	1.46178	15.3	12	8.15	645.7	7.1	0.115847
8	8.45	717	82.5	0.140366	7	1.76133	18.4	14.5	8.45	716.4	7.2	0.138966
9	8.65	768	88.4	0.100887	5	0.917869	9.6	7.6	8.65	768.2	5.1	0.10108
10	9.3	819	94.2	0.0343268	7	0.374217	3.9	3.1	9.3	817.2	7.3	0.0287598

PeakCounter: 10

**DISPLAY GEL LANE LETTER:** Lane b
**DIGITAL REPORT GEL LANE:** 3
**LANE TYPE:** Specimen
**ACTUAL GEL LANE:** 3
Go to Gel Schematic
Return to Lane Navigator
Band	pI	Loc	% RF	Amplitude	Width	Integr. OD	% Integr. OD	% Lane IntOD	pI	G. Loc	G. Sigma	G. Ampl
1	4.06	18	2.1	0.069921	7	0.841743	5.8	5	4.06	17.9	6.6	0.0717464
2	4.57	96	11	0.0210294	6	0.237298	1.6	1.4	4.57	96.9	6.2	0.0216418
3	4.733	121	13.9	0.0590265	6	0.570998	4	3.4	4.733	120.6	6.2	0.0520198
4	4.857	140	16.1	0.155736	11	3.00027	20.8	17.9	4.857	139.3	10.9	0.154627
5	4.981	159	18.2	0.0729645	9	1.11623	7.7	6.6	4.981	160.8	9.2	0.0684114
6	5.422	229	26.3	0.006852	5	0.0507495	0.4	0.3	5.422	226.5	5.2	0.0054823
7	5.55	250	28.7	0.0669803	6	0.716958	5	4.3	5.55	247.4	6	0.0671205
8	5.623	262	30	0	1	0	0	0	5.623	235	1	0
9	5.708	276	31.7	0.193816	4	1.45011	10.1	8.6	5.708	275.3	4.1	0.201268
10	5.787	289	33.1	0.125281	4	0.882717	6.1	5.3	5.787	289.1	4.2	0.118365
Band	pI	Loc	% RF	Amplitude	Width	Integr. OD	% Integr. OD	% Lane IntOD	pI	G. Loc	G. Sigma	G. Ampl
11	5.824	295	33.8	0.113733	3	0.384054	2.7	2.3	5.824	295.7	3	0.0716561
12	5.865	302	34.6	0.0357749	8	0.598589	4.2	3.6	5.865	300.6	8.5	0.039765
13	6.07	338	38.8	0.0572887	6	0.56783	3.9	3.4	6.07	337.3	5.9	0.0545183
14	6.138	350	40.1	0.181114	4	1.34448	9.3	8	6.138	349.5	4.1	0.185904
15	6.257	371	42.5	0.0737201	9	1.16797	8.1	7	6.257	369	9.4	0.0702233
16	6.337	385	44.2	0.0418945	2	0.0794232	0.6	0.5	6.337	385.5	1.6	0.02827
17	6.462	407	46.7	0.120672	6	1.32805	9.2	7.9	6.462	406.2	6.1	0.122979
18	6.625	431	49.4	0.0087141	4	0.0743442	0.5	0.4	6.625	432.3	4.3	0.0098457

PeakCounter: 18

**DISPLAY GEL LANE LETTER:** Lane c
**DIGITAL REPORT GEL LANE:** 4
**LANE TYPE:** Co-Migrated
**ACTUAL GEL LANE:** 4
Go to Gel Schematic
Return to Lane Navigator
Band	pI	Loc	% RF	Amplitude	Width	Integr. OD	% Integr. OD	% Lane IntOD	pI	G. Loc	G. Sigma	G. Ampl
1	4.069	22	2.5	0.0706395	5	0.707231	3.4	3.3	4.069	21.2	5.5	0.072733
2	4.55	96	11.1	0.0545237	6	0.412704	2	1.9	4.55	94.7	6	0.0391043
3	4.752	127	14.6	0.132346	17	3.67406	17.4	17	4.752	120.1	17.3	0.120081
4	4.843	141	16.2	0.175198	12	3.17697	15.1	14.7	4.843	143	11.9	0.150102
5	4.998	165	19	0.0630248	10	1.09642	5.2	5.1	4.998	162.1	10.1	0.0610554
6	5.2	196	22.6	0.0312765	5	0.278034	1.3	1.3	5.2	195	5.3	0.0298561
7	5.444	233	26.8	0.0113644	3	0.0590322	0.3	0.3	5.444	232.5	3.1	0.0106846
8	5.576	253	29.1	0.0920139	4	0.584731	2.8	2.7	5.576	252	3.7	0.0894639
9	5.648	264	30.4	0.0056474	5	0.284079	1.3	1.3	5.648	271.7	4.8	0.0332349
10	5.74	278	32	0.253735	4	1.46889	7	6.8	5.74	277.2	3.7	0.224359
Band	pI	Loc	% RF	Amplitude	Width	Integr. OD	% Integr. OD	% Lane IntOD	pI	G. Loc	G. Sigma	G. Ampl
11	5.813	289	33.3	0.164208	5	1.33962	6.3	6.2	5.813	289	5.1	0.148079
12	5.857	296	34.1	0.139575	4	0.788892	3.7	3.7	5.857	296.8	4	0.11179
13	5.918	307	35.4	0.0297672	6	0.402824	1.9	1.9	5.918	304.2	5.7	0.0396484
14	6.094	339	39.1	0.035926	6	0.308928	1.5	1.4	6.094	339.9	5.8	0.0300356
15	6.171	353	40.7	0.151232	3	0.918414	4.4	4.3	6.171	352.6	3.5	0.148466
16	6.286	374	43.1	0.0634449	7	0.808646	3.8	3.7	6.286	373.4	7.2	0.0630878
17	6.369	389	44.8	0.0354918	2	0.0968985	0.5	0.4	6.369	389.3	1.7	0.0322278
18	6.49	411	47.4	0.112136	7	1.41747	6.7	6.6	6.49	410.5	7.4	0.108035
19	6.55	422	48.6	0.0221023	2	0.076172	0.4	0.4	6.55	420.4	2.2	0.0193
20	6.677	436	50.2	0.006449	3	0.0299559	0.1	0.1	6.677	435.1	2.9	0.0058083
Band	pI	Loc	% RF	Amplitude	Width	Integr. OD	% Integr. OD	% Lane IntOD	pI	G. Loc	G. Sigma	G. Ampl
21	6.85	455	52.4	0.0215841	4	0.156684	0.7	0.7	6.85	455.6	4.2	0.0208854
22	7.35	515	59.3	0.0336858	5	0.299485	1.4	1.4	7.35	514.6	5.2	0.0323721
23	8.15	640	73.7	0.0438746	15	1.58073	7.5	7.3	8.15	628.4	14.6	0.061118
24	8.45	717	82.6	0.0569935	5	0.532915	2.5	2.5	8.45	716.4	5.4	0.0556366
25	8.65	765	88.1	0.0481587	4	0.324597	1.5	1.5	8.65	764.5	3.8	0.0477081
26	9.3	816	94	0.017568	12	0.279196	1.3	1.3	9.3	814.8	11.6	0.0135826

PeakCounter: 26

**DISPLAY GEL LANE LETTER:** Lane d
**DIGITAL REPORT GEL LANE:** 5
**LANE TYPE:** pI Markers
**ACTUAL GEL LANE:** 5
Go to Gel Schematic
Return to Lane Navigator
Band	pI	Loc	% RF	Amplitude	Width	Integr. OD	% Integr. OD	% Lane IntOD	pI	G. Loc	G. Sigma	G. Ampl
1	4.55	89	10.2	0.0680114	4	0.483998	6.5	4.8	4.55	88.7	4	0.0683993
2	5.2	177	20.3	0.062167	5	0.610545	8.2	6.1	5.2	177.3	5.5	0.0626656
3	5.85	291	33.4	0.0789925	4	0.561345	7.6	5.6	5.85	290.9	4	0.0790634
4	6.55	415	47.7	0.140619	7	1.56979	21.2	15.6	6.55	414.6	6.6	0.134088
5	6.85	451	51.8	0.0352107	5	0.279971	3.8	2.8	6.85	451.9	4.9	0.0319806
6	7.35	515	59.2	0.0544306	5	0.447069	6	4.4	7.35	514.2	5	0.0503752
7	8.15	641	73.7	0.108141	6	1.17844	15.9	11.7	8.15	640.6	6.4	0.103395
8	8.45	712	81.8	0.101763	7	1.31003	17.7	13	8.45	711.5	7	0.105933
9	8.65	762	87.6	0.0959252	4	0.733278	9.9	7.3	8.65	762.2	4.3	0.0966223
10	9.3	816	93.8	0.0263401	6	0.243315	3.3	2.4	9.3	815.6	5.8	0.0238424

PeakCounter: 10

**DISPLAY GEL LANE LETTER:** Lane e
**DIGITAL REPORT GEL LANE:** 6
**LANE TYPE:** Specimen
**ACTUAL GEL LANE:** 6
Go to Gel Schematic
Return to Lane Navigator
Band	pI	Loc	% RF	Amplitude	Width	Integr. OD	% Integr. OD	% Lane IntOD	pI	G. Loc	G. Sigma	G. Ampl
1	4.052	21	2.4	0.0938241	5	0.873713	4.1	3.6	4.052	21.5	5.1	0.0958439
2	4.615	99	11.4	0.0383595	4	0.126838	0.6	0.5	4.615	97	3.9	0.0184766
3	4.752	118	13.6	0.148917	15	4.03973	18.8	16.5	4.752	119.7	15.4	0.147792
4	4.954	146	16.8	0.218232	12	4.22269	19.7	17.3	4.954	145.4	11.6	0.205262
5	5.092	165	19	0.112342	9	1.56219	7.3	6.4	5.092	165	9	0.0975421
6	5.459	226	26	0.0168988	3	0.0751463	0.3	0.3	5.459	225.8	2.6	0.016155
7	5.571	246	28.3	0.166699	3	0.787266	3.7	3.2	5.571	245.7	2.7	0.16723
8	5.656	261	30	0.0190097	4	0.120698	0.6	0.5	5.656	261.4	3.5	0.0193298
9	5.735	275	31.6	0.287166	3	1.69061	7.9	6.9	5.735	275	3.3	0.285197
10	5.819	290	33.4	0.145809	3	0.679695	3.2	2.8	5.819	288.7	3.2	0.119876
Band	pI	Loc	% RF	Amplitude	Width	Integr. OD	% Integr. OD	% Lane IntOD	pI	G. Loc	G. Sigma	G. Ampl
11	5.853	296	34.1	0.136198	4	0.683335	3.2	2.8	5.853	295.3	4.2	0.0921542
12	5.905	305	35.1	0.0567257	16	1.48005	6.9	6	5.905	303	15.6	0.0536598
13	6.077	335	38.6	0.0630213	7	0.70082	3.3	2.9	6.077	335.4	6.8	0.0580273
14	6.145	347	39.9	0.219147	3	1.18109	5.5	4.8	6.145	346.6	3.2	0.210453
15	6.237	363	41.8	0.0365138	2	0.0941696	0.4	0.4	6.237	360.2	2.3	0.0230551
16	6.277	370	42.6	0.0825274	6	0.944238	4.4	3.9	6.277	369.4	6.3	0.0841269
17	6.369	386	44.4	0.0720706	3	0.288839	1.3	1.2	6.369	386.7	2.5	0.0648154
18	6.507	410	47.2	0.145764	8	1.88707	8.8	7.7	6.507	407.5	7.7	0.138441
19	6.681	433	49.8	0.0094984	3	0.0397746	0.2	0.2	6.681	432.9	2.5	0.0088186

PeakCounter: 19

**DISPLAY GEL LANE LETTER:** Lane f
**DIGITAL REPORT GEL LANE:** 7
**LANE TYPE:** pI Markers
**ACTUAL GEL LANE:** 7
Go to Gel Schematic
Return to Lane Navigator
Band	pI	Loc	% RF	Amplitude	Width	Integr. OD	% Integr. OD	% Lane IntOD	pI	G. Loc	G. Sigma	G. Ampl
1	4.55	90	10.4	0.0775353	4	0.524351	7.3	5.3	4.55	90.3	3.8	0.078148
2	5.2	182	21	0.08712	4	0.624381	8.7	6.3	5.2	181.6	4.2	0.0847912
3	5.85	299	34.5	0.0871486	3	0.460837	6.4	4.7	5.85	299.3	2.9	0.0882692
4	6.55	419	48.3	0.122735	5	1.13772	15.9	11.5	6.55	418.3	5.4	0.118589
5	6.85	454	52.4	0.0387143	5	0.305265	4.3	3.1	6.85	454	4.5	0.0382036
6	7.35	519	59.9	0.0627022	5	0.509587	7.1	5.2	7.35	518.3	4.7	0.0608857
7	8.15	640	73.8	0.104205	6	1.172	16.4	11.8	8.15	640	6.5	0.102298
8	8.45	714	82.4	0.127124	6	1.28526	17.9	13	8.45	713.6	5.7	0.126597
9	8.65	768	88.6	0.107425	4	0.744397	10.4	7.5	8.65	767.5	4	0.106295
10	9.3	822	94.8	0.0304695	8	0.398826	5.6	4	9.3	820.8	8.2	0.0272826

PeakCounter: 10

Gel Banding Patterns Display in HTML using GIF stacking technique
(Uses Migration Position Normalized Data from above text peak reports)
Return to Lane Navigator Go to Gel Schematic Key

a b c d e f

MK SP CO MK SP MK

pI 4.55 label for gel schematic

pI 5.20 label for gel schematic

pI 5.8 label for gel schematic

pI 6.55 label for gel schematic

pI 6.85 label for gel schematic

pI 7.35 label for gel schematic

pI 8.15 label for gel schematic

pI 8.45 label for gel schematic

pI 8.65 label for gel schematic

pI 9.30 label for gel schematic

pI 4.55 label for gel schematic

pI 5.20 label for gel schematic

pI 5.8 label for gel schematic

pI 6.55 label for gel schematic

pI 6.85 label for gel schematic

pI 7.35 label for gel schematic

pI 8.15 label for gel schematic

pI 8.45 label for gel schematic

pI 8.65 label for gel schematic

pI 9.30 label for gel schematic

Key to Gel Schematic:

MK=pI Marker Lanes, pI bands are in red
SP=Specimen Lanes, Specimen bands are in blue
CO=Co-migrated Lanes of pI Markers and Specimen (pI bands are in red, Specimen bands are in blue)
Note: In co-migrated lanes, some pI Markers and Specimen bands may overlap in experiment. Red pI
band is reported when overlapped peak position is equal to nominal pI Marker position. Blue Specimen
band is reported when overlapped peak position deviates slightly from nominal pI Marker position.
Weak (<2% rel. intensity), Medium (2-6% rel. intensity), and Strong (>6% rel. intensity) band intensities indicated by gel schematic band thickness.
Return to Top of Gel Schematic

PeakCount: 93

HTML Forms TextArea Normalized Peak-Position Peak-Area for Copy-Paste events
4.550 9.600 5.200 9.100 5.850 7.300 6.550 14.900 6.850 4.300 7.350 7.500 8.150 15.300 8.450 18.400 8.650 9.600 9.300 3.900 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0	DISPLAY GEL LANE LETTER: Lane a DIGITAL REPORT GEL LANE: 2 LANE TYPE: pI Markers ACTUAL GEL LANE: 2 PeakCount: 10 Return to Lane Navigator
4.060 5.800 4.570 1.600 4.733 4.000 4.857 20.800 4.981 7.700 5.422 0.400 5.550 5.000 5.623 0 5.708 10.100 5.787 6.100 5.824 2.700 5.865 4.200 6.070 3.900 6.138 9.300 6.257 8.100 6.337 0.600 6.462 9.200 6.625 0.500 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0	DISPLAY GEL LANE LETTER: Lane b DIGITAL REPORT GEL LANE: 3 LANE TYPE: Specimen ACTUAL GEL LANE: 3 PeakCount: 18 Return to Lane Navigator
4.069 3.400 4.550 2.000 4.752 17.400 4.843 15.100 4.998 5.200 5.200 1.300 5.444 0.300 5.576 2.800 5.648 1.300 5.740 7.000 5.813 6.300 5.857 3.700 5.918 1.900 6.094 1.500 6.171 4.400 6.286 3.800 6.369 0.500 6.490 6.700 6.550 0.400 6.677 0.100 6.850 0.700 7.350 1.400 8.150 7.500 8.450 2.500 8.650 1.500 9.300 1.300 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0	DISPLAY GEL LANE LETTER: Lane c DIGITAL REPORT GEL LANE: 4 LANE TYPE: Co-Migrated ACTUAL GEL LANE: 4 PeakCount: 26 Return to Lane Navigator
4.550 6.500 5.200 8.200 5.850 7.600 6.550 21.200 6.850 3.800 7.350 6.000 8.150 15.900 8.450 17.700 8.650 9.900 9.300 3.300 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0	DISPLAY GEL LANE LETTER: Lane d DIGITAL REPORT GEL LANE: 5 LANE TYPE: pI Markers ACTUAL GEL LANE: 5 PeakCount: 10 Return to Lane Navigator
4.052 4.100 4.615 0.600 4.752 18.800 4.954 19.700 5.092 7.300 5.459 0.300 5.571 3.700 5.656 0.600 5.735 7.900 5.819 3.200 5.853 3.200 5.905 6.900 6.077 3.300 6.145 5.500 6.237 0.400 6.277 4.400 6.369 1.300 6.507 8.800 6.681 0.200 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0	DISPLAY GEL LANE LETTER: Lane e DIGITAL REPORT GEL LANE: 6 LANE TYPE: Specimen ACTUAL GEL LANE: 6 PeakCount: 19 Return to Lane Navigator
4.550 7.300 5.200 8.700 5.850 6.400 6.550 15.900 6.850 4.300 7.350 7.100 8.150 16.400 8.450 17.900 8.650 10.400 9.300 5.600 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0	DISPLAY GEL LANE LETTER: Lane f DIGITAL REPORT GEL LANE: 7 LANE TYPE: pI Markers ACTUAL GEL LANE: 7 PeakCount: 10 Return to Lane Navigator

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EXPERIMENTAL CONDITIONS: Gels were scanned at 600 d.p.i. in Adobe Photoshop 3.02 using a Sharp JX-325 Scanner with Film Scanner Attachment. Gels were analyzed for Densitometry, Intregration, and Migration Position Normalization in EXCEL 5.0 Format. *.xls files were imported into Paradox 7.0 to save as *.db files.

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Delphi 4.0 Generation of this Isoelectric Focusing (IEF) RFE PAGE 5 Tools in HTML
This HTML is automatically generated from a series of Paradox 7.0 database filesof IEF peak reports is controlled with a main RFEIEF Paradox 7.0 database withDelphi 3.0 front end (see Delphi 2.0 form) and back end for facile addition and modification of the Content of these RFE PAGE 5 HTML tools.
Contact Brad Tenge at btenge@ora.fda.gov for details.

RFE Contacts
A summary list of RFE contact personnel is available at http://www.cfsan.fda.gov/~frf/rfecon.html.

For isoelectric focusing laboratory experiments for fish species identification, you may contact

Brad Tenge at E-mail: btenge@ora.fda.gov
Ngoc-Lan Nguyen at E-mail: nnguyen@ora.fda.gov
Jim Barnett at E-mail: jbarnett@ora.fda.gov
Pat Rogers at E-mail: pxr@cfsan.fda.gov

For isoelectric focusing gel analysis and pattern recognition applications, you may contact

Brad Tenge at E-mail: btenge@ora.fda.gov
Ngoc-Lan Nguyen at E-mail: nnguyen@ora.fda.gov
Fred Fry at E-mail frf@vm.cfsan.fda.gov

For CFSAN web consultation and posting, you may contact

Fred Fry E-mail: frf@vm.cfsan.fda.gov

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Last updated on 2003-FEB-27 by bjt/frf/nln

Peak Report Data from Isoelectric Focusing for a specimen of: Orange Roughy ---- Go to Display Gel Lane Navigator.

Peak Report Data from Isoelectric Focusing for a specimen of:
Orange Roughy ---- Go to Display Gel Lane Navigator.