Goat anti-Rabbit IgG (Heavy & Light Chain) Antibody (FITC)

Details for Product No. ABIN101988, Supplier: Log in to see
Antigen
Epitope
Heavy & Light Chain
6235
2031
1776
1135
782
648
372
305
34
34
31
15
12
12
12
12
11
7
3
3
1
1
1
1
1
1
1
Reactivity
Rabbit
3081
2914
2380
1662
1509
786
737
489
476
411
401
395
342
298
271
160
159
111
55
34
30
26
16
15
12
10
8
7
5
5
4
4
3
3
3
3
2
2
2
2
2
2
1
1
1
1
1
1
1
Host
Goat
7133
5286
1973
1077
507
362
88
84
51
35
23
15
8
5
3
1
1
Clonality
Polyclonal
Conjugate
FITC
2227
1822
1751
1266
794
718
430
343
235
147
145
140
140
116
113
112
102
98
94
93
92
91
89
84
81
80
78
75
73
72
68
68
65
61
55
54
54
54
54
53
53
53
53
53
53
52
48
43
41
37
32
31
23
23
22
20
19
18
17
16
15
15
15
14
14
14
12
12
12
12
12
11
11
10
10
10
10
10
9
9
9
9
9
8
8
8
8
8
7
6
6
6
6
6
6
6
5
5
5
5
5
5
5
5
4
4
4
4
3
3
3
3
3
3
3
3
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
1
1
1
1
1
1
Application
FLISA, Immunomicroscopy (IM), Western Blotting (WB)
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'Independent Validation' Badge
Antigen Rabbit IgG (Heavy & Light Chain)
Lot Number 611-1202
Method validated Immunofluorescence
Positive Control Lab stock CBD-SNAP antibody
Negative Control No SNAP-tag antibody
Notes We validate the specificity of the secondary goat anti-rabbit IgG (heavy & light chain) antibody (FITC) ABIN101988 for rabbit IgG antibody.
Primary Antibody ABIN1573927
Secondary Antibody ABIN101988
Protocol
  • 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.
Experimental Notes 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.
Validation Images
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
Isotype IgG
Specificity IgG (H&L)
Characteristics Concentration Definition: by UV absorbance at 280 nm
Antigen
Application Notes 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.
Comment

Excitation/Emission wavelength: 494 nm/514 nm

Restrictions For Research Use only
Format Lyophilized
Reconstitution Restore with deionized water (or equivalent)
Concentration 2.0 mg/mL
Buffer 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Preservative Sodium azide
Precaution of Use 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.
Handling Advice Product is photosensitive and should be protected from light.
Supplier Images
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 ...
Product cited in: Jiang, Xia, Yang, Shao, Liao, Zhu, Jiang: "Novel insights into a treatment for aplastic anemia based on the advanced proliferation of bone marrow‑derived mesenchymal stem cells induced by fibroblast growth factor 1." in: Molecular medicine reports, Vol. 12, Issue 6, pp. 7877-82, 2016 (PubMed).

Background publications 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).