Ziege anti-Kaninchen IgG (Heavy & Light Chain) Antikörper (FITC)

Details zu Produkt Nr. ABIN101988, Anbieter: Anmelden zum Anzeigen
Antigen
Epitop
Heavy & Light Chain
6301
1957
1804
1065
889
727
375
301
35
34
31
15
11
7
3
3
1
1
1
1
1
1
1
Reaktivität
Kaninchen
3073
2887
2376
1634
1481
785
690
459
456
397
369
364
301
288
280
146
128
114
42
34
29
26
16
15
11
10
8
7
5
5
4
4
3
3
3
3
3
3
3
2
2
2
1
1
1
1
1
1
1
Wirt
Ziege
6986
5101
1927
1048
535
361
90
84
55
35
23
15
8
5
3
1
1
Klonalität
Polyklonal
Konjugat
FITC
2225
1808
1765
1261
763
687
455
310
196
155
155
150
127
122
122
102
102
96
94
89
84
79
78
75
73
68
68
68
65
61
55
55
54
54
54
53
53
53
53
53
53
53
48
48
41
38
35
34
31
24
23
23
22
20
18
18
17
16
15
15
15
14
14
14
14
14
12
12
12
12
12
10
10
10
10
10
9
9
9
9
8
8
8
8
8
7
6
6
6
6
6
5
5
5
5
5
5
4
4
4
4
4
4
4
4
4
4
3
3
3
3
3
3
3
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
1
1
1
1
1
1
1
Applikation
FLISA, Immunomicroscopy (IM), Western Blotting (WB)
Optionen
Hersteller
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Hersteller Produkt- Nr.
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'Independent Validation' Siegel
Antigen Rabbit IgG (Heavy & Light Chain)
Chargennummer 611-1202
Validierte Anwendung Immunofluorescence
Positivkontrolle Lab stock CBD-SNAP antibody
Negativkontrolle No SNAP-tag antibody
Bewertung We validate the specificity of the secondary goat anti-rabbit IgG (heavy & light chain) antibody (FITC) ABIN101988 for rabbit IgG antibody.
Primärantikörper ABIN1573927
Sekundärantikörper ABIN101988
Protokoll
  • Oyster visceral mass tissue is dissected and fixed in 4% paraformaldehyde in seawater overnight.
  • Serial dehydration process using an automated ASP300S Enclosed Tissue Processor (Leica Biosystems) as follows:
    • 70% ethanol for 45min
    • 90% ethanol for 45min
    • 90% ethanol for 45min
    • 100% ethanol twice for 45min
    • xylene twice for 45min
    • paraffin wax at 58°C 3 times for 30 min
  • Tissue is mounted in a paraffin block and hardened overnight before.
  • 8µm tissue sections are retrieved from the block and collected on circular glass cover slips.
  • Heat cover slips at 60°C for 1h.
  • Deparaffination and rehydration:
    • Xylene twice for 15min
    • 100% ethanol twice for 10min
    • 95% ethanol for 10 min
    • 85% ethanol for 10 min
    • 70% ethanol for 10 min
    • 50% ethanol for 10 min
    • 30% ethanol for 10 min
    • distilled water for 10 min
    • PBS for 10 min
  • Wash tissue sections with PBS with 0.05% triton X twice for 30min.
  • Permeabilize in PBS with 0.05% triton X overnight.
  • Treatment of the tissue sections with 1mg/mL sodium borohydride in PBS three times for 5min to reduce autofluorescence.
  • Wash sections in PBS 3 times for 15 min for at RT.
  • Block sections in PBST with 1% BSA for 2 hours at RT.
  • Incubate sections with CBD-SNAP antibody (lab stock) diluted 1:200 in PBST with 1% BSA overnight at 4°C to detect the location of chitin.
  • Wash sections in PBS 3 times for 15min with PBS at RT.
  • Additionally, incubate the CBD-SNAP and SNAP-tag double-stained sections with rabbit anti-SNAP antibody (antibodies-online, ABIN1573927, lot 13D000621) diluted 1:200 in PBST with 1% BSA overnight at 4°C.
  • Wash sections in PBS 3 times for 15min with PBS at RT.
  • Incubate sections with the secondary goat anti-rabbit IgG (heavy & light chain) antibody (FITC) (antibodies-online, ABIN101988, lot 611-1202) diluted 1:400 in PBST with 1% BSA for 2h at °C.
  • Wash sections in PBS three times for 15min at RT.
  • Counterstain with 0.1µg/mL DAPI in PBS for 15min at RT.
  • Wash sections in PBS three times for 15min at RT.
  • Mount sections on a microscopic slide using 50% glycerol in PBS.
  • Seal cover slips with nail polish.
  • Confocal imaging on Leica SPE.
  • Visualization of the data performed on LAS 3D software.
Anmerkungen To validate the specificity of the anti-rabbit FITC secondary antibody ABIN101988, 8µm paraffin sections of oyster visceral mass were observed in this study. We compared the fluorescence signals with immunofluorescence study. The negative control specimen was always compared with the test specimen or the positive control specimen on the same day, using the same laser power, gain, offset, accumulation/averaging settings on the Leica SPE confocal microscope. Visualization of the data was performed on LAS 3D software, with the same visualization setting to compare signal brightness. We found that the samples treated with anti-rabbit FITC secondary antibody ABIN101988 had similar fluorescence signals as the positive control Anti Rabbit Alexa 488. Excitation at the same laser wavelength and power did not generate fluorescence in the negative control section, when anti-rabbit FITC secondary antibody was applied in the absence of rabbit produced anti-SNAP antibody.
Validierungsbilder
Immunofluorescence validation image for Goat anti-Rabbit IgG (Heavy & Light Chain) antibody (FITC) (ABIN101988) Immunofluorescence images of oyster visceral mass tissue, with the specificity of Ant...
Immunogen Rabbit IgG whole molecule
Isotyp IgG
Spezifität IgG (H&L)
Produktmerkmale Concentration Definition: by UV absorbance at 280 nm
Antigen
Applikations-hinweise This product is designed for immunofluorescence microscopy, fluorescence based plate assays (FLISA) and fluorescent western blotting. This product is also suitable for multiplex analysis, including multicolor imaging, utilizing various commercial platforms.
Kommentare

Excitation/Emission wavelength: 494 nm/514 nm

Beschränkungen Nur für Forschungszwecke einsetzbar
Format Lyophilized
Rekonstitution Restore with deionized water (or equivalent)
Konzentration 2.0 mg/mL
Buffer 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Konservierungs-mittel Sodium azide
Vorsichtsmaßnahmen WARNING: Reagents contain sodium azide. Sodium azide is very toxic if ingested or inhaled. Avoid contact with skin, eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid under acidic conditions. Dilute azide-containing compounds in running water before discarding to avoid accumulation of potentially explosive deposits in lead or copper plumbing.
Handhabung Product is photosensitive and should be protected from light.
Bilder des Herstellers
Western Blotting (WB) image for Goat anti-Rabbit IgG (Heavy & Light Chain) antibody (FITC) (ABIN101988) FITC (fluorescein) and HRP (horse radish peroxidase) conjugated secondary antibody wa...
Dot Blot (DB) image for Goat anti-Rabbit IgG (Heavy & Light Chain) antibody (FITC) (ABIN101988) FITC (fluorescein) and HRP (horse radish peroxidase) conjugated secondary antibody wa...
Western Blotting (WB) image for Goat anti-Rabbit IgG (Heavy & Light Chain) antibody (FITC) (ABIN101988) Western Blot of Anti-Rabbit IgG (H&L) (GOAT) Antibody . Lane M: 3 μl Molecular Ladder...
Western Blotting (WB) image for Goat anti-Rabbit IgG (Heavy & Light Chain) antibody (FITC) (ABIN101988) Western blot of Fluorescein conjugated Goat Anti-Rabbit IgG secondary antibody. Lane ...
Allgemeine Veröffentlichungen Blakeslee, Baines: "Immunofluorescence using dichlorotriazinylaminofluorescein (DTAF). I. Preparation and fractionation of labelled IgG." in: Journal of immunological methods, Vol. 13, Issue 3-4, pp. 305-20, 1977 (PubMed).