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: 19950801gel01
Unique Web Code: org01Srr
Tissue Aliquot Code: hoplatla I94-001 #11a Gel Analysis Conditions


Gel Image	Fish Image	Calibrated Gel	Comparison 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:** 6
**LANE TYPE:** pI Markers
**ACTUAL GEL LANE:** 8
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	3.5	41	3.7	0.158882	18	5.05746	13.9	11.2	3.5	44.3	17.9	0.159642
2	4.55	181	16.2	0.278465	6	2.69899	7.4	6	4.55	181.6	5.8	0.261138
3	5.2	261	23.3	0.449422	5	3.87762	10.6	8.6	5.2	261.3	5	0.436348
4	5.85	410	36.6	0.653465	3	3.85002	10.6	8.6	5.85	409.7	3.3	0.662948
5	6.55	569	50.8	0.377936	5	3.31656	9.1	7.4	6.55	568.7	5.2	0.359564
6	6.85	607	54.2	0.125646	7	1.46811	4	3.3	6.85	607.9	7.1	0.116256
7	7.35	697	62.3	0.294314	7	3.27881	9	7.3	7.35	695.7	6.6	0.279361
8	8.15	858	76.7	0.125783	8	1.71347	4.7	3.8	8.15	857.5	7.8	0.124163
9	8.45	918	82	0.174911	6	1.87461	5.1	4.2	8.45	918	6	0.176321
10	8.65	957	85.5	0.124517	6	1.20388	3.3	2.7	8.65	956.3	5.6	0.121255
Band	pI	Loc	% RF	Amplitude	Width	Integr. OD	% Integr. OD	% Lane IntOD	pI	G. Loc	G. Sigma	G. Ampl
11	9.3	1085	97	0.570744	9	8.09289	22.2	18	9.3	1083.8	8.6	0.531841

PeakCounter: 11

**DISPLAY GEL LANE LETTER:** Lane b
**DIGITAL REPORT GEL LANE:** 7
**LANE TYPE:** Specimen
**ACTUAL GEL LANE:** 9
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	3.808	88	7.8	0.243164	5	2.26048	4.9	4	3.808	88	5.3	0.238556
2	4.066	121	10.8	0.486732	5	4.57693	9.8	8.2	4.066	121.5	5.3	0.490855
3	4.175	135	12	0.496795	6	5.55356	11.9	9.9	4.175	135	6.5	0.484571
4	4.511	178	15.9	0.640412	7	7.77549	16.7	13.9	4.511	177.2	6.7	0.658447
5	5.534	341	30.4	0.287901	8	3.48381	7.5	6.2	5.534	342.8	7.9	0.249904
6	5.629	363	32.4	0.773123	6	7.35524	15.8	13.1	5.629	362.1	6.3	0.660276
7	5.668	372	33.2	0.48639	9	7.83342	16.8	14	5.668	375.6	9.4	0.468459
8	6.032	455	40.6	0.256774	3	1.24643	2.7	2.2	6.032	454.9	2.8	0.249011
9	6.094	469	41.8	0.142364	2	0.368387	0.8	0.7	6.094	468.7	2	0.102135
10	6.196	492	43.9	0.221837	16	5.52817	11.9	9.9	6.196	488	15.7	0.199212
Band	pI	Loc	% RF	Amplitude	Width	Integr. OD	% Integr. OD	% Lane IntOD	pI	G. Loc	G. Sigma	G. Ampl
11	6.346	526	46.9	0.134599	2	0.517088	1.1	0.9	6.346	526.7	2.3	0.127127

PeakCounter: 11

**DISPLAY GEL LANE LETTER:** Lane c
**DIGITAL REPORT GEL LANE:** 8
**LANE TYPE:** pI Markers
**ACTUAL GEL LANE:** 10
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	3.5	53	4.7	0.189995	19	6.579	16.3	13.5	3.5	52.2	19.2	0.193149
2	4.55	182	16.2	0.345925	5	3.03143	7.5	6.2	4.55	182.3	5.4	0.314922
3	5.2	264	23.5	0.53191	5	4.20452	10.4	8.6	5.2	263.9	4.6	0.515755
4	5.85	415	37	0.635381	4	4.29369	10.7	8.8	5.85	414.7	3.7	0.653268
5	6.55	572	50.9	0.405146	5	3.39393	8.4	6.9	6.55	571.8	4.9	0.389146
6	6.85	609	54.2	0.13318	7	1.45658	3.6	3	6.85	609.2	6.8	0.121073
7	7.35	699	62.2	0.355284	6	3.70663	9.2	7.6	7.35	698.1	6.2	0.338606
8	8.15	859	76.5	0.124298	8	1.686	4.2	3.5	8.15	858.3	7.7	0.123886
9	8.45	918	81.7	0.187947	6	1.858	4.6	3.8	8.45	917.4	5.7	0.182754
10	8.65	955	85	0.151958	6	1.484	3.7	3	8.65	955	5.7	0.147657
Band	pI	Loc	% RF	Amplitude	Width	Integr. OD	% Integr. OD	% Lane IntOD	pI	G. Loc	G. Sigma	G. Ampl
11	9.3	1084	96.5	0.60301	9	8.57673	21.3	17.6	9.3	1082.2	8.6	0.565257

PeakCounter: 11

**DISPLAY GEL LANE LETTER:** Lane d
**DIGITAL REPORT GEL LANE:** 9
**LANE TYPE:** Co-Migrated
**ACTUAL GEL LANE:** 11
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	3.5	57	5.1	0.103785	18	3.25729	5.1	4.4	3.5	58.7	18	0.101837
2	3.795	93	8.3	0.289691	5	2.54272	4	3.4	3.795	93	5	0.289197
3	4.074	127	11.3	0.494486	5	4.54294	7.1	6.1	4.074	127.2	5.1	0.501989
4	4.189	141	12.5	0.541916	7	6.28456	9.9	8.4	4.189	141.2	6.6	0.533276
5	4.55	185	16.5	0.637761	7	7.77955	12.2	10.5	4.55	185.5	6.7	0.658263
6	5.2	274	24.4	0.250803	8	3.25708	5.1	4.4	5.2	273.4	8.3	0.222141
7	5.514	344	30.6	0.228393	2	1.01103	1.6	1.4	5.514	344.6	2.4	0.242317
8	5.612	366	32.6	0.71724	4	3.46889	5.5	4.7	5.612	366.1	3.6	0.537445
9	5.666	378	33.6	0.353861	12	7.24282	11.4	9.7	5.666	376	11.9	0.344275
10	5.85	419	37.3	0.3602	4	2.33138	3.7	3.1	5.85	419.4	3.6	0.363564
Band	pI	Loc	% RF	Amplitude	Width	Integr. OD	% Integr. OD	% Lane IntOD	pI	G. Loc	G. Sigma	G. Ampl
11	6.047	461	41	0.265057	3	1.18256	1.9	1.6	6.047	461.2	2.5	0.263317
12	6.113	475	42.3	0.171768	5	0.897323	1.4	1.2	6.113	476.1	4.5	0.111941
13	6.221	498	44.3	0.26341	13	5.21116	8.2	7	6.221	494.9	13.2	0.222578
14	6.386	533	47.4	0.170166	3	0.752886	1.2	1	6.386	533	2.8	0.154248
15	6.55	568	50.5	0.157776	7	1.71301	2.7	2.3	6.55	566.6	6.9	0.140934
16	6.85	606	53.9	0.108442	7	1.22351	1.9	1.6	6.85	606.8	6.8	0.100819
17	7.35	696	61.9	0.239879	6	2.47828	3.9	3.3	7.35	694.7	6	0.231477
18	8.15	852	75.8	0.00387122	1	0	0	0	8.15	839	1	0
19	8.45	913	81.2	0.131364	7	1.49815	2.4	2	8.45	913.6	6.6	0.128914
20	8.65	955	85	0.123866	6	1.21962	1.9	1.6	8.65	954.2	5.7	0.120955
Band	pI	Loc	% RF	Amplitude	Width	Integr. OD	% Integr. OD	% Lane IntOD	pI	G. Loc	G. Sigma	G. Ampl
21	9.3	1081	96.2	0.46168	7	5.67808	8.9	7.6	9.3	1080.1	7.5	0.429742

PeakCounter: 21

**DISPLAY GEL LANE LETTER:** Lane e
**DIGITAL REPORT GEL LANE:** 10
**LANE TYPE:** Specimen
**ACTUAL GEL LANE:** 12
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	3.801	91	8.1	0.150351	5	1.3557	4.5	3.7	3.801	90.9	5.2	0.148106
2	4.077	124	11	0.555457	5	4.41723	14.6	12	4.077	124.3	4.6	0.542067
3	4.186	137	12.2	0.394245	5	3.49855	11.6	9.5	4.186	137.6	5.3	0.373937
4	4.554	181	16.1	0.664225	6	6.68925	22.2	18.2	4.554	181.3	5.5	0.683442
5	5.524	339	30.2	0.194212	2	0.802279	2.7	2.2	5.524	339.2	2.3	0.197393
6	5.62	361	32.1	0.782873	4	5.24551	17.4	14.3	5.62	361.5	3.9	0.76032
7	5.664	371	33	0.418422	5	3.52317	11.7	9.6	5.664	371.3	5.2	0.383696
8	6.035	454	40.4	0.176886	3	0.893678	3	2.4	6.035	453.5	3	0.166891
9	6.094	467	41.5	0.0806403	2	0.260384	0.9	0.7	6.094	467.3	2.3	0.0641086
10	6.199	490	43.6	0.150108	13	3.09226	10.2	8.4	6.199	486.1	13.5	0.129263
Band	pI	Loc	% RF	Amplitude	Width	Integr. OD	% Integr. OD	% Lane IntOD	pI	G. Loc	G. Sigma	G. Ampl
11	6.354	524	46.6	0.0877397	3	0.412543	1.4	1.1	6.354	524.6	3	0.0763515

PeakCounter: 11

**DISPLAY GEL LANE LETTER:** Lane f
**DIGITAL REPORT GEL LANE:** 11
**LANE TYPE:** pI Markers
**ACTUAL GEL LANE:** 13
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	3.5	54	4.8	0.178481	18	5.83808	16	12.9	3.5	53.8	18.3	0.179759
2	4.55	177	15.7	0.30613	5	2.57029	7	5.7	4.55	177.1	5.2	0.278389
3	5.2	257	22.8	0.474858	5	3.77182	10.3	8.3	5.2	256.8	4.6	0.45927
4	5.85	409	36.3	0.582349	4	3.90918	10.7	8.6	5.85	409.4	3.7	0.59361
5	6.55	567	50.4	0.357164	5	3.02	8.3	6.7	6.55	566.5	5	0.339242
6	6.85	604	53.6	0.138962	7	1.52731	4.2	3.4	6.85	604.9	6.8	0.126245
7	7.35	695	61.7	0.280461	7	3.16826	8.7	7	7.35	693.9	6.8	0.263719
8	8.15	856	76	0.106785	9	1.6412	4.5	3.6	8.15	854.6	8.8	0.104642
9	8.45	914	81.2	0.156388	6	1.71415	4.7	3.8	8.45	913.1	6.3	0.153778
10	8.65	952	84.5	0.110589	6	1.18242	3.2	2.6	8.65	951	6.2	0.107921
Band	pI	Loc	% RF	Amplitude	Width	Integr. OD	% Integr. OD	% Lane IntOD	pI	G. Loc	G. Sigma	G. Ampl
11	9.3	1078	95.7	0.592785	8	8.19332	22.4	18.1	9.3	1076.5	8.4	0.550334

PeakCounter: 11

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 3.50 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

pI 3.50 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: 76

HTML Forms TextArea Normalized Peak-Position Peak-Area for Copy-Paste events
3.500 13.900 4.550 7.400 5.200 10.600 5.850 10.600 6.550 9.100 6.850 4.000 7.350 9.000 8.150 4.700 8.450 5.100 8.650 3.300 9.300 22.200 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 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: 6 LANE TYPE: pI Markers ACTUAL GEL LANE: 8 PeakCount: 11 Return to Lane Navigator
3.808 4.900 4.066 9.800 4.175 11.900 4.511 16.700 5.534 7.500 5.629 15.800 5.668 16.800 6.032 2.700 6.094 0.800 6.196 11.900 6.346 1.100 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 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: 7 LANE TYPE: Specimen ACTUAL GEL LANE: 9 PeakCount: 11 Return to Lane Navigator
3.500 16.300 4.550 7.500 5.200 10.400 5.850 10.700 6.550 8.400 6.850 3.600 7.350 9.200 8.150 4.200 8.450 4.600 8.650 3.700 9.300 21.300 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	DISPLAY GEL LANE LETTER: Lane c DIGITAL REPORT GEL LANE: 8 LANE TYPE: pI Markers ACTUAL GEL LANE: 10 PeakCount: 11 Return to Lane Navigator
3.500 5.100 3.795 4.000 4.074 7.100 4.189 9.900 4.550 12.200 5.200 5.100 5.514 1.600 5.612 5.500 5.666 11.400 5.850 3.700 6.047 1.900 6.113 1.400 6.221 8.200 6.386 1.200 6.550 2.700 6.850 1.900 7.350 3.900 8.150 0 8.450 2.400 8.650 1.900 9.300 8.900 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	DISPLAY GEL LANE LETTER: Lane d DIGITAL REPORT GEL LANE: 9 LANE TYPE: Co-Migrated ACTUAL GEL LANE: 11 PeakCount: 21 Return to Lane Navigator
3.801 4.500 4.077 14.600 4.186 11.600 4.554 22.200 5.524 2.700 5.620 17.400 5.664 11.700 6.035 3.000 6.094 0.900 6.199 10.200 6.354 1.400 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	DISPLAY GEL LANE LETTER: Lane e DIGITAL REPORT GEL LANE: 10 LANE TYPE: Specimen ACTUAL GEL LANE: 12 PeakCount: 11 Return to Lane Navigator
3.500 16.000 4.550 7.000 5.200 10.300 5.850 10.700 6.550 8.300 6.850 4.200 7.350 8.700 8.150 4.500 8.450 4.700 8.650 3.200 9.300 22.400 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	DISPLAY GEL LANE LETTER: Lane f DIGITAL REPORT GEL LANE: 11 LANE TYPE: pI Markers ACTUAL GEL LANE: 13 PeakCount: 11 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: 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.