Lapin anti-Chévre IgG (Whole Molecule) Anticorps (Colloidal Gold (5nm))

Détails pour le produit réf. ABIN3042038, Fournisseur: Connectez-vous pour afficher
Antigène
Épitope
Whole Molecule
5028
2577
1931
1069
1007
857
330
299
35
34
31
8
7
7
3
3
2
2
1
1
1
1
1
1
Reactivité
Chévre
2912
2859
2405
1752
1484
796
623
442
381
380
355
340
307
293
208
148
139
114
41
29
27
25
12
11
9
8
6
5
5
5
4
4
4
4
4
4
4
4
3
3
3
2
2
1
1
1
1
1
Hôte
Lapin
6893
4911
1878
895
703
401
92
54
52
21
9
8
5
5
2
1
1
Clonalité
Polyclonal
Conjugué
Colloidal Gold (5nm)
2235
1889
1740
1374
765
679
519
300
299
288
274
268
217
188
178
124
103
79
69
60
54
46
43
40
40
36
34
32
29
29
27
27
27
27
27
27
27
27
22
22
19
19
19
19
16
15
15
15
15
15
14
12
12
11
10
10
9
9
9
9
9
9
9
8
8
8
8
7
7
6
6
6
6
6
6
5
5
5
5
5
4
4
4
4
4
3
3
3
3
3
3
2
2
2
2
2
2
2
2
2
2
2
2
1
1
1
1
1
1
1
1
Application
Immunoelectron Microscopy (IEM), Immunocytochemistry (ICC), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))
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'Independent Validation' signe
Antigène Rabbit IgG
Application validée Electron Microscopy
Contrôle positif Lab stock CBD-SNAP antibody
Contrôle négative No SNAP-tag antibody
Conclusion We validate that the anti-Rabbit IgG (Whole Molecule) Antibody (Colloidal Gold (5nm)) ABIN3042038 specifically a rabbit primary antibody in SEM-EBSD images of oyster visceral mass tissue.
Anticorps primaire ABIN1573927
Anticorps secondaire ABIN3042038
Protocole
  • Oyster visceral mass tissue is dissected and fixed in 4% paraformaldehyde in seawater overnight.
  • Sections are rinsed with 70% ethanol and then transferred to 70% ethanol for about 2h at RT before automated dehydration process.
  • Serial dehydration process using an automated ASP300S Enclosed Tissue Processor (Leica Biosystems) as follows:
    • 70% ethanol for 45min
    • 90% ethanol twice for 45min
    • 100% ethanol twice for 45min
    • xylene twice for 45min
    • paraffin wax at 58°C 3 times for 30min
  • Tissue is mounted in a paraffin block and hardened overnight.
  • 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 10min
    • 85% ethanol for 10min
    • 70% ethanol for 10min
    • 50% ethanol for 10min
    • 30% ethanol for 10min
    • distilled water for 10min
    • PBS for 10min
  • 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 15min for at RT.
  • Block sections in PBST with 1% BSA for 2h 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 at RT.
  • Incubate sections with the secondary rabbit anti-rabbit IgG (whole molecule) antibody Colloidal Gold (5nm) conjugate (antibodies-online, ABIN3042038) diluted 1:50 in PBST with 1% BSA in the dark overnicht at 4°C.
  • Wash sections in PBS three times for 15min at RT.
  • Post fixsections with 1% glutaraldehyde in PBS for 15min at RT.
  • Wash sections in distilled water three times for 10min at RT.
  • R-Gent SE-EM silver enhancement reagents (Aurion, 2551, lot 60323/2):
    • Prepare a slower developer mix with 1:60 ratio of initiator:activator.
    • Silver enhancement for 30 min with 1:20 developer: enhancer ratio.
    • Stop the reaction by washing with distilled water for 5min 3 times.
  • Graded alcohol series: /li>
    • 50% alcohol for 15min
    • 70% alcohol for 15min
    • 85% alcohol for 15min
    • 95% alcohol for 15min
    • 100% alcohol for 15min
  • Critical point drying of the sections with CO2 transitional fluid for 15min.
  • Slowly degas sections at a degassing rate of 100 Pa/min.
  • Sections are mounted on the same aluminum stub, coated with carbon and visualized at 20kV with probe current at 40, with basic SEM-EBSD mode on a Hitachi S-3400 Variable Pressure SEM, working distance kept at 10mm. Brightness and contrast settings are kept constant for the both samples.
Notes To validate the specificity of the anti-Rabbit IgG (Whole Molecule) Antibody (Colloidal Gold (5nm)) ABIN3042038, 8µm paraffin sections of oyster’s visceral mass were observed in this study. We compared the level of silver aggregating ability in the presence and absence of ABIN3042038, using commercially available silver enhancement method and SEM-EBSD imaging. We found that the test samples treated with anti-Rabbit IgG (Whole Molecule) Antibody (Colloidal Gold (5nm)) showed distinctive silver aggregates while the primary rabbit anti-SNAP antibody ABIN1573927 was present. These aggregates were not observable in the negative control.
Validation Images
Electron Microscopy validation image for Rabbit anti-Goat IgG (Whole Molecule) antibody (Colloidal Gold (5nm)) (ABIN3042038) SEM-EBSD images of oyster visceral mass tissue stained with CBD-SNAP and anti-SNAP pr...
Immunogène Goat IgG (whole molecule)
Isotype IgG
Specificité This antibody is specific for goat IgG
Aucune reactivité croisée Humain, Rat (Rattus), Souris, Lapin
Réactivité croisée (Details) This colloidal gold conjugated antibody is specific for goat IgG and shows no cross-reactivity with human/ rat/mouse/rabbit IgG.
Purification This antibody is purified from antiserum by immunoaffinity chromatography which removes essentially all rabbit serum proteins, except the specific antibody for goat IgG. The rabbit anti-goat IgG is conjugated to 5 nm colloidal gold and generates electron dense particles that are visible using electron microscopy.
Domaine de recherche Immunology, Secondary Antibodies
Indications d'application Electron Microscopy|1:20-50| Immunohistochemistry(Paraffin-embedded Section)|1:50-100| Immunohistochemistry(Frozen Section)|1:50-100| Immunocytochemistry|1:50-100|
Restrictions For Research Use only
Format Liquid
Conseil sur la manipulation Do not freeze.
Stock 4 °C
Stockage commentaire At 4°C for one year.
Date de péremption 12 months