Chèvre anti-Lapin IgG (Heavy & Light Chain) Anticorps (FITC)

Détails pour le produit réf. ABIN101988, Fournisseur: Connectez-vous pour afficher
Antigène
Épitope
Heavy & Light Chain
4683
2573
1882
1039
1015
825
305
273
35
34
31
8
7
7
3
3
2
2
1
1
1
1
1
1
Reactivité
Lapin
2780
2760
2346
1724
1436
778
607
426
364
338
330
324
307
290
169
148
126
114
41
29
27
25
12
11
9
8
6
5
5
4
4
4
4
4
4
4
4
3
3
3
2
2
2
1
1
1
1
1
Hôte
Chèvre
6749
4661
1834
812
684
394
88
52
51
9
8
5
5
4
2
1
1
Clonalité
Polyclonal
Conjugué
FITC
2183
1861
1718
1362
765
675
521
297
295
284
274
268
217
186
178
117
102
79
69
58
54
40
35
26
23
22
19
19
19
19
19
16
15
15
15
15
15
15
14
13
12
11
11
10
10
9
9
9
9
9
9
9
8
8
8
7
6
6
6
6
6
6
6
5
5
5
5
5
4
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
1
1
1
1
1
1
1
1
Application
FLISA, Immunomicroscopy (IM), Western Blotting (WB)
Options
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Images

'Independent Validation' signe
Antigène Rabbit IgG (Heavy & Light Chain)
Numéro du lot 611-1202
Application validée Immunofluorescence
Contrôle positif Lab stock CBD-SNAP antibody
Contrôle négative No SNAP-tag antibody
Conclusion We validate the specificity of the secondary goat anti-rabbit IgG (heavy & light chain) antibody (FITC) ABIN101988 for rabbit IgG antibody.
Anticorps primaire ABIN1573927
Anticorps secondaire ABIN101988
Protocole
  • 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.
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...
Immunogène Rabbit IgG whole molecule
Isotype IgG
Specificité IgG (H&L)
Attributs du produit Concentration Definition: by UV absorbance at 280 nm
Domaine de recherche Immunology, Secondary Antibodies
Indications d'application 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.
Commentaires

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
Agent conservateur Sodium azide
Précaution d'utilisation 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.
Conseil sur la manipulation Product is photosensitive and should be protected from light.
Supplier Images
Western Blotting (WB) image for Chèvre anti-Lapin IgG (Heavy & Light Chain) Anticorps (FITC) (ABIN101988) FITC (fluorescein) and HRP (horse radish peroxidase) conjugated secondary antibody wa...
Background publications Blakeslee, Baines: "Immunofluorescence using dichlorotriazinylaminofluorescein (DTAF). I. Preparation and fractionation of labelled IgG." dans: Journal of immunological methods, Vol. 13, Issue 3-4, pp. 305-20, 1977 (PubMed).