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:
Blackback Flounder ---- Go to Display Gel Lane Navigator.

FDA Seafood List Scientific Name: Pleuronectes americanus
FDA Seafood List Common Name: Blackback Flounder
FDA Seafood List Market Name: Flounder (Sole)
Unique IEF Gel Code: 19980331gel01
Unique Web Code: bbh01Srr
Tissue Aliquot Code: pleuamer H94-002 #08m 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 pI Markers	Lane d Co-Migrated	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	27	3.4	0.105852	3	0.640257	7	5.1	4.55	27.1	3.5	0.104026
2	5.2	126	16	0.0989963	5	0.808427	8.9	6.4	5.2	126.6	4.8	0.0951706
3	5.85	235	29.9	0.0572064	4	0.429878	4.7	3.4	5.85	234.6	4.3	0.055798
4	6.55	379	48.2	0.154096	6	1.61612	17.7	12.9	6.55	378.3	6.4	0.143094
5	6.85	415	52.8	0.040483	5	0.329347	3.6	2.6	6.85	415.5	5.3	0.0349579
6	7.35	485	61.7	0.0942581	4	0.711227	7.8	5.7	7.35	484.7	4.2	0.0952113
7	8.15	609	77.5	0.10196	8	1.463	16	11.6	8.15	608.8	8.3	0.0994717
8	8.45	670	85.2	0.137906	7	1.70532	18.7	13.6	8.45	669.1	7.1	0.136031
9	8.65	716	91.1	0.115936	5	0.946482	10.4	7.5	8.65	715.1	4.7	0.11312
10	9.3	762	96.9	0.0582751	5	0.481889	5.3	3.8	9.3	761.2	5	0.0549208

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	4.446	18	2.3	0.0192342	4	0.142708	0.5	0.4	4.446	17.9	4.1	0.0194866
2	4.609	43	5.4	0.216949	18	6.11358	21.3	16.3	4.609	53.6	18	0.191627
3	4.972	99	12.5	0.0401002	18	1.2315	4.3	3.3	4.972	86.8	18	0.0386006
4	5.324	155	19.5	0.160621	5	1.38522	4.8	3.7	5.324	156.4	5.1	0.152896
5	5.353	160	20.2	0.155521	5	0.533488	1.9	1.4	5.353	162.3	4.9	0.0610752
6	5.488	183	23.1	0.231012	5	2.11249	7.4	5.6	5.488	183.9	5.2	0.230426
7	5.618	205	25.9	0.219864	6	1.86501	6.5	5	5.618	206.1	5.8	0.181694
8	5.718	222	28	0.11942	14	3.03836	10.6	8.1	5.718	219.5	14.1	0.121823
9	5.872	249	31.4	0.01943	12	0.423753	1.5	1.1	5.872	242.8	12.5	0.0191453
10	5.952	265	33.4	0.0129519	4	0.0621709	0.2	0.2	5.952	266.7	4.4	0.00792147
Band	pI	Loc	% RF	Amplitude	Width	Integr. OD	% Integr. OD	% Lane IntOD	pI	G. Loc	G. Sigma	G. Ampl
11	6.122	299	37.7	0.309506	5	2.83792	9.9	7.6	6.122	299.3	5.1	0.316752
12	6.181	311	39.2	0.2894	5	2.50129	8.7	6.7	6.181	311.2	5.4	0.259423
13	6.251	325	41	0.0661267	9	1.0303	3.6	2.7	6.251	325.7	9	0.0643454
14	6.326	340	42.9	0.0837048	6	0.804776	2.8	2.1	6.326	339.8	5.7	0.0801858
15	6.391	353	44.5	0.126748	3	0.666554	2.3	1.8	6.391	354.1	3.1	0.122551
16	6.485	372	46.9	0.0123513	3	0.0779415	0.3	0.2	6.485	372.4	3.4	0.0128275
17	6.618	393	49.6	0.0626514	4	0.415157	1.4	1.1	6.618	393.2	3.8	0.0617285
18	7.307	484	61	0.169058	5	1.40822	4.9	3.7	7.307	484.2	4.9	0.161087
19	7.466	508	64.1	0.0939499	14	2.05273	7.2	5.5	7.466	508.5	13.8	0.0841984

PeakCounter: 19

**DISPLAY GEL LANE LETTER:** Lane c
**DIGITAL REPORT GEL LANE:** 4
**LANE TYPE:** pI Markers
**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	4.55	26	3.3	0.0881221	4	0.687011	7.1	5.2	4.55	26	4.3	0.0898501
2	5.2	127	16.2	0.0608118	7	0.748897	7.7	5.7	5.2	124.8	7	0.0605351
3	5.85	239	30.4	0.0415	6	0.420855	4.4	3.2	5.85	239.1	5.9	0.0404235
4	6.55	376	47.9	0.140879	7	1.65051	17.1	12.5	6.55	375.1	7.2	0.128794
5	6.85	411	52.4	0.0457391	5	0.378468	3.9	2.9	6.85	410.7	4.8	0.0444194
6	7.35	480	61.1	0.0991602	4	0.77341	8	5.9	7.35	479.4	4.4	0.0986023
7	8.15	604	76.9	0.106044	8	1.48527	15.4	11.3	8.15	603.3	8.4	0.100028
8	8.45	667	85	0.146659	7	1.81846	18.8	13.8	8.45	666.6	7.2	0.142887
9	8.65	718	91.5	0.11936	5	0.96964	10	7.4	8.65	717.3	4.7	0.115885
10	9.3	761	96.9	0.0827213	5	0.738958	7.6	5.6	9.3	760.4	5.3	0.078266

PeakCounter: 10

**DISPLAY GEL LANE LETTER:** Lane d
**DIGITAL REPORT GEL LANE:** 5
**LANE TYPE:** Co-Migrated
**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	4.464	15	1.9	0.0270869	7	0.419951	2.1	1.5	4.464	18.7	7.1	0.0333846
2	4.55	27	3.4	0.0603861	4	0.37275	1.8	1.3	4.55	27.6	4	0.0524758
3	4.636	39	5	0.125894	6	1.34184	6.7	4.7	4.636	40	6.2	0.121602
4	5.2	118	15	0.0369665	6	0.378451	1.9	1.3	5.2	118.1	5.7	0.0375001
5	5.345	145	18.5	0.063364	6	0.224266	1.1	0.8	5.345	145.4	5.9	0.0214923
6	5.383	152	19.4	0.0479408	7	0.723028	3.6	2.5	5.383	148.7	6.9	0.0588821
7	5.522	178	22.7	0.130176	6	1.48968	7.4	5.2	5.522	175.6	6.2	0.13615
8	5.635	199	25.4	0.17897	6	1.84791	9.2	6.4	5.635	198.9	6	0.174926
9	5.721	215	27.4	0.0919552	11	1.70617	8.5	5.9	5.721	215.6	10.9	0.0885642
10	5.968	261	33.2	0.00924425	4	0.0788735	0.4	0.3	5.968	260.9	4.4	0.010185
Band	pI	Loc	% RF	Amplitude	Width	Integr. OD	% Integr. OD	% Lane IntOD	pI	G. Loc	G. Sigma	G. Ampl
11	6.139	293	37.3	0.302315	4	2.31204	11.5	8	6.139	292.8	4.3	0.301435
12	6.197	304	38.7	0.226048	3	1.40179	7	4.9	6.197	303.5	3.5	0.228163
13	6.294	322	41	0.0703586	5	0.635533	3.2	2.2	6.294	320.7	5.3	0.0673706
14	6.352	333	42.4	0.0754021	7	0.902631	4.5	3.1	6.352	332.6	6.9	0.0741504
15	6.427	347	44.2	0.123034	3	0.715425	3.5	2.5	6.427	347.2	3.4	0.117191
16	6.55	370	47.1	0.0327842	6	0.343976	1.7	1.2	6.55	369.8	5.8	0.0337402
17	6.692	388	49.4	0.0373944	3	0.180833	0.9	0.6	6.692	387.7	2.7	0.0380055
18	6.85	408	52	0.0239949	7	0.255396	1.3	0.9	6.85	408.6	6.8	0.0211521
19	7.298	474	60.4	0.150217	5	1.23565	6.1	4.3	7.298	473.8	5.1	0.136286
20	7.439	495	63.1	0.0759857	8	0.97393	4.8	3.4	7.439	495.8	7.8	0.0703384
Band	pI	Loc	% RF	Amplitude	Width	Integr. OD	% Integr. OD	% Lane IntOD	pI	G. Loc	G. Sigma	G. Ampl
21	8.15	602	76.7	0.0507777	9	0.807757	4	2.8	8.15	601.7	9.4	0.0484425
22	8.45	666	84.8	0.0792601	6	0.811353	4	2.8	8.45	665.6	5.8	0.0786484
23	8.65	713	90.8	0.069229	4	0.509616	2.5	1.8	8.65	712.9	4.3	0.0661701
24	9.3	761	96.9	0.0508926	6	0.491035	2.4	1.7	9.3	761.1	5.7	0.0484529

PeakCounter: 24

**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	4.498	28	3.6	0.0630082	8	0.819483	2.9	2.3	4.498	27.6	7.7	0.0601573
2	4.658	51	6.5	0.345584	12	4.92098	17.6	14.1	4.658	55	11.9	0.233193
3	5.029	104	13.2	0.0499927	18	3.00298	10.8	8.6	5.029	78	18	0.0941267
4	5.383	161	20.4	0.174379	3	0.940108	3.4	2.7	5.383	160.1	3.2	0.163386
5	5.417	167	21.2	0.105447	4	0.757297	2.7	2.2	5.417	166.1	4	0.106609
6	5.552	191	24.2	0.300733	4	2.10185	7.5	6	5.552	191.3	3.8	0.309328
7	5.692	216	27.4	0.321231	5	2.56078	9.2	7.3	5.692	215.7	4.5	0.317921
8	5.782	232	29.4	0.119284	9	1.87249	6.7	5.4	5.782	232	9.4	0.112874
9	6.004	274	34.8	0.0236291	3	0.127442	0.5	0.4	6.004	274.4	3.1	0.0230134
10	6.142	301	38.2	0.434378	4	2.8549	10.2	8.2	6.142	301.3	3.7	0.430329
Band	pI	Loc	% RF	Amplitude	Width	Integr. OD	% Integr. OD	% Lane IntOD	pI	G. Loc	G. Sigma	G. Ampl
11	6.204	313	39.7	0.376161	3	1.87577	6.7	5.4	6.204	312.4	2.8	0.374286
12	6.286	329	41.8	0.0789855	10	1.28396	4.6	3.7	6.286	328.4	10	0.0724371
13	6.347	341	43.3	0.0880428	5	0.664314	2.4	1.9	6.347	342.4	4.8	0.0773588
14	6.429	357	45.3	0.129802	3	0.707413	2.5	2	6.429	357.1	3.1	0.129019
15	6.532	377	47.8	0.0128039	3	0.055547	0.2	0.2	6.532	377.1	2.5	0.0123579
16	6.691	397	50.4	0.0551116	3	0.309796	1.1	0.9	6.691	396.9	3.2	0.0543849
17	7.307	482	61.2	0.154277	5	1.28723	4.6	3.7	7.307	481.9	5.2	0.139032
18	7.447	503	63.8	0.0901772	11	1.7673	6.3	5.1	7.447	503.4	11.4	0.0871617

PeakCounter: 18

**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	30	3.9	0.0975074	4	0.690778	7	5.1	4.55	29.6	4	0.0982569
2	5.2	125	16	0.0642566	7	0.772083	7.8	5.7	5.2	125.6	6.7	0.0650196
3	5.85	235	30.2	0.0645037	4	0.437812	4.4	3.3	5.85	235.2	4.1	0.0608731
4	6.55	377	48.4	0.16963	6	1.64502	16.7	12.2	6.55	376.6	5.9	0.157079
5	6.85	409	52.5	0.0536384	5	0.443289	4.5	3.3	6.85	409.4	5.1	0.0493983
6	7.35	481	61.7	0.103235	4	0.705259	7.2	5.2	7.35	481.3	3.9	0.102336
7	8.15	602	77.3	0.0890843	10	1.4809	15	11	8.15	602.6	9.8	0.0850247
8	8.45	669	85.9	0.145548	8	1.97784	20.1	14.7	8.45	667.4	7.8	0.143229
9	8.65	716	91.9	0.129227	4	0.964994	9.8	7.2	8.65	715.4	4.4	0.124489
10	9.3	762	97.8	0.0755294	6	0.729719	7.4	5.4	9.3	760.6	6.1	0.0671746

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 MK CO 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: 91

HTML Forms TextArea Normalized Peak-Position Peak-Area for Copy-Paste events
4.550 7.000 5.200 8.900 5.850 4.700 6.550 17.700 6.850 3.600 7.350 7.800 8.150 16.000 8.450 18.700 8.650 10.400 9.300 5.300 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 a DIGITAL REPORT GEL LANE: 2 LANE TYPE: pI Markers ACTUAL GEL LANE: 2 PeakCount: 10 Return to Lane Navigator
4.446 0.500 4.609 21.300 4.972 4.300 5.324 4.800 5.353 1.900 5.488 7.400 5.618 6.500 5.718 10.600 5.872 1.500 5.952 0.200 6.122 9.900 6.181 8.700 6.251 3.600 6.326 2.800 6.391 2.300 6.485 0.300 6.618 1.400 7.307 4.900 7.466 7.200 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 b DIGITAL REPORT GEL LANE: 3 LANE TYPE: Specimen ACTUAL GEL LANE: 3 PeakCount: 19 Return to Lane Navigator
4.550 7.100 5.200 7.700 5.850 4.400 6.550 17.100 6.850 3.900 7.350 8.000 8.150 15.400 8.450 18.800 8.650 10.000 9.300 7.600 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 c DIGITAL REPORT GEL LANE: 4 LANE TYPE: pI Markers ACTUAL GEL LANE: 4 PeakCount: 10 Return to Lane Navigator
4.464 2.100 4.550 1.800 4.636 6.700 5.200 1.900 5.345 1.100 5.383 3.600 5.522 7.400 5.635 9.200 5.721 8.500 5.968 0.400 6.139 11.500 6.197 7.000 6.294 3.200 6.352 4.500 6.427 3.500 6.550 1.700 6.692 0.900 6.850 1.300 7.298 6.100 7.439 4.800 8.150 4.000 8.450 4.000 8.650 2.500 9.300 2.400 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 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: Co-Migrated ACTUAL GEL LANE: 5 PeakCount: 24 Return to Lane Navigator
4.498 2.900 4.658 17.600 5.029 10.800 5.383 3.400 5.417 2.700 5.552 7.500 5.692 9.200 5.782 6.700 6.004 0.500 6.142 10.200 6.204 6.700 6.286 4.600 6.347 2.400 6.429 2.500 6.532 0.200 6.691 1.100 7.307 4.600 7.447 6.300 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	DISPLAY GEL LANE LETTER: Lane e DIGITAL REPORT GEL LANE: 6 LANE TYPE: Specimen ACTUAL GEL LANE: 6 PeakCount: 18 Return to Lane Navigator
4.550 7.000 5.200 7.800 5.850 4.400 6.550 16.700 6.850 4.500 7.350 7.200 8.150 15.000 8.450 20.100 8.650 9.800 9.300 7.400 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 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

Thank you for using the Regulatory Fish Encyclopedia (RFE)!

RFE Species Browse | RFE Tutorial | Seafoods | CFSAN

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

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

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