Kaninchen anti-Ziege IgG (Whole Molecule) Antikörper (Colloidal Gold (5nm))

Details zu Produkt Nr. ABIN3042038, Anbieter: Anmelden zum Anzeigen
Antigen
Epitop
Whole Molecule
5120
1885
1841
1055
1039
866
330
304
35
34
31
12
8
7
3
3
2
1
1
1
1
1
1
1
Reaktivität
Ziege
2914
2617
2172
1556
1392
751
601
456
426
389
365
357
295
280
265
145
132
113
41
30
29
26
16
11
10
8
7
6
5
5
4
4
4
3
3
3
3
3
3
2
2
2
1
1
1
1
1
Wirt
Kaninchen
6502
4929
1660
1009
521
357
93
67
54
19
8
5
5
5
3
1
1
Klonalität
Polyklonal
Konjugat
Colloidal Gold (5nm)
2222
1825
1740
1284
772
695
447
307
219
171
165
141
137
129
110
107
77
75
71
65
65
56
55
54
54
53
53
53
53
53
53
52
50
40
37
35
24
19
18
18
17
15
15
15
14
14
14
14
12
12
11
10
10
10
10
10
9
9
9
9
9
8
8
8
8
6
6
6
6
6
6
5
5
5
5
5
4
4
4
4
4
4
4
4
4
4
3
3
3
3
3
3
2
2
2
2
2
2
2
2
2
2
2
2
2
2
1
1
1
1
1
1
Applikation
Immunoelectron Microscopy (IEM), Immunocytochemistry (ICC), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))
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'Independent Validation' Siegel
Antigen Rabbit IgG
Validierte Anwendung Electron Microscopy
Positivkontrolle Lab stock CBD-SNAP antibody
Negativkontrolle No SNAP-tag antibody
Bewertung 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.
Primärantikörper ABIN1573927
Sekundärantikörper ABIN3042038
Protokoll
  • 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.
Anmerkungen 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.
Validierungsbilder
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...
Immunogen Goat IgG (whole molecule)
Isotyp IgG
Spezifität This antibody is specific for goat IgG
Keine Kreuzreaktivität Human, Ratte (Rattus), Maus, Kaninchen
Kreuzreaktivität (Details) This colloidal gold conjugated antibody is specific for goat IgG and shows no cross-reactivity with human/ rat/mouse/rabbit IgG.
Reinigung 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.
Applikations-hinweise Electron Microscopy|1:20-50| Immunohistochemistry(Paraffin-embedded Section)|1:50-100| Immunohistochemistry(Frozen Section)|1:50-100| Immunocytochemistry|1:50-100|
Beschränkungen Nur für Forschungszwecke einsetzbar
Format Liquid
Handhabung Do not freeze.
Lagerung 4 °C
Informationen zur Lagerung At 4°C for one year.
Haltbarkeit 12 months