Âne anti-Chévre IgG (Heavy & Light Chain) Anticorps (IRDye680RD)

Détails pour le produit réf. ABIN2169629, 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
Chévre
2780
2760
2347
1724
1435
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
Âne
6750
4661
1833
812
684
394
88
52
51
9
8
5
5
4
2
1
1
Clonalité
Polyclonal
Conjugué
IRDye680RD
2183
1862
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
8
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
In-Gel Western blotting (gelWB), Immunohistochemistry (IHC), Western Blotting (WB)
Options
Fournisseur
Connectez-vous pour afficher
Supplier Product No.
Connectez-vous pour afficher

Images

'Independent Validation' signe
Antigène Goat IgG (Heavy & Light Chain)
Numéro du lot C50330-03
Application validée Western Blotting
Contrôle positif Human A375 and SKMEL 28 melanoma cell line lysates; C57BL/6 mouse serum
Anticorps primaire ABIN1440014
Anticorps secondaire Donkey anti-Goat IgG (H+L) IRDye680RD (LiCor, antibodies-online product number ABIN2169630 , lot number: C50330-03)
Protocole
  • WB on HeLa cells and human melanoma cell lines SKMEL28 and A375
    • Mouse serum was collected from C57BL/6 mice and lysed in 6M Urea/20mM Tris.
    • Exosomes were extracted with ExoQuick reagent (SBI, product #EXOQ5A, lot #140624-001) following the manufacturer’s protocol and resuspended in 30µl of modified RIPA buffer.
    • 20µl, 10µl, and 5µl aliquots of 5-fold diluted exosome solution were used.
    • Samples were denatured in Bio-Rad Laemmli sample buffer (product # 1610737, lot #t350001755)containing beta-marcaptoethanol and separated on 4-20% Bio-Rad TGX gel (product #456-1034, lot #t64041496).
    • Transfer onto PVDF membrane (Millipore, product #IPLF00010, lot #R5EA5898E) was done on TransBlot SD apparatus (Bio-Rad) following manufacturer’s protocol.
    • Block membranes with LiCor Blocking Buffer (product 927-40000, lot V1381) for 2h.
    • Incubation with
      • primary CD63 antibody ABIN1440014 at 4°C diluted 1:500 in LiCor Blocking Buffer at 4°C overnight.
      • primary rabbit anti-CD9 antibody (Proteintech, 20597-1-AP, lot 00013334) diluted 1:500 in LiCor Blocking Buffer at 4°C overnight.
    • Wash 4x 15min with PBS-T.
    • Incubation with secondary antibody LiCor Donkey anti-Goat IgG (Heavy & Light Chain) Antibody (IRDye680RD) secondary antibody (antibodies-online product number ABIN2169630, lot C50330-03, 1:15000 dilution) and incubated on a shaker for 1h at RT.
    • Three rinses plus four 15min washes—2 PBS-T, 2 PBS (as required by LiCor).
    • Blot was developed on a LiCor imaging system (Odyssey 9120, scan resolution: 169um, image quality: low).
  • WB on C57BL/6 mouse serum exosomes
    • Mouse serum was collected from C57BL/6 mice and lysed in 6M Urea/20mM Tris.
    • Exosomes were extracted with ExoQuick reagent (SBI, product #EXOQ5A, lot #140624-001) following the manufacturer’s protocol and resuspended in 30µl of modified RIPA buffer.
    • 20µl, 10µl, and 5µl aliquots of 5-fold diluted exosome solution were used.
    • Samples were denatured in Bio-Rad Laemmli sample buffer (product # 1610737, lot #t350001755)containing beta-marcaptoethanol and separated on 4-20% Bio-Rad TGX gel (product #456-1034, lot #t64041496).
    • Transfer onto PVDF membrane (Millipore, product #IPLF00010, lot #R5EA5898E) was done on TransBlot SD apparatus (Bio-Rad) following manufacturer’s protocol.
    • Block membranes with LiCor Blocking Buffer (product 927-40000, lot V1381) for 2h.
    • Incubation with
      • primary CD63 antibody ABIN1440014 at 4°C diluted 1:500 in LiCor Blocking Buffer at 4°C overnight.
      • primary rabbit anti-CD9 antibody (Proteintech, 20597-1-AP, lot 00013334) diluted 1:500 in LiCor Blocking Buffer at 4°C overnight.
    • Wash 4x 15min with PBS-T.
    • Incubation with secondary antibody LiCor Donkey anti-Goat IgG (Heavy & Light Chain) Antibody (IRDye680RD) secondary antibody (antibodies-online, ABIN2169630, lot C50330-03, 1:15000 dilution) and incubated on a shaker for 1h at RT.
    • Three rinses plus four 15min washes—2 PBS-T, 2 PBS (as required by LiCor).
    • Blot was developed on a LiCor imaging system (Odyssey 9120, scan resolution: 169um, image quality: low).
Notes The donkey anti-Goat IgG (H&L) IRDye680RD-conjugated secondary antibody ABIN2169630 works successfully in Western blot to reveal goat IgG pirmary antibodies used on human cell lysate and prepared mouse serum exosome samples.
Validation Images
Western Blotting validation image for Donkey anti-Goat IgG (Heavy & Light Chain) antibody (IRDye680RD) (ABIN2169630) A. 20µg total protein of cultured HeLa cells and human melanoma cell lines SKMEL28 an...
Marque In-Cell Western™,CellTag™
Immunogène Goat IgG
Isotype IgG
Fragment Whole molecule
Specificité Goat IgG
Based on ELISA, this antibody reacts with the heavy and light chains of goat IgG and with sheep IgG.
 Réactivité croisée Mouton
Réactivité croisée (Details) This antibody was tested by Dot Blot and/or solid-phase adsorbed for minimal cross-reactivity with human, mouse, rabbit, rat, chicken, guinea pig, hamster, swine, and horse serum proteins, but may cross-react with immunoglobulins from other species.
Attributs du produit Blocking buffers and antibody diluents made with bovine serum albumin (BSA) and dry milk may contain IgG that reacts with anti-bovine IgG, anti-goat IgG, anti-horse IgG, and anti-sheep IgG antibodies. This can lead to a significant increase in background and/or reduction of secondary antibody titer for protein detection applications.
Purification immunoaffinity chromatography
Domaine de recherche Immunology, Secondary Antibodies
Indications d'application Western blot: 1:5,000 - 1:25,000
Restrictions For Research Use only
Format Lyophilized
Reconstitution Reconstitute with sterile, distilled water
Concentration 1.0 mg/mL
Buffer PBS, pH 7.4
Agent conservateur Sodium azide
Précaution d'utilisation This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Conseil sur la manipulation Protect from light.
Stock 4 °C
Produit citée dans: Alayev, Berger, Kramer, Schwartz, Holz: "The combination of rapamycin and resveratrol blocks autophagy and induces apoptosis in breast cancer cells." dans: Journal of cellular biochemistry, Vol. 116, Issue 3, pp. 450-7, 2015 (PubMed).