Ziege anti-Kaninchen IgG (Heavy & Light Chain) Antikörper (IRDye680LT)

Details zu Produkt Nr. ABIN2169666, Anbieter: Anmelden zum Anzeigen
Antigen
Epitop
Heavy & Light Chain
6301
1957
1804
1065
889
727
375
301
35
34
31
15
11
7
3
3
1
1
1
1
1
1
1
Reaktivität
Kaninchen
3073
2887
2376
1634
1481
785
690
459
456
397
369
364
301
288
280
146
128
114
42
34
29
26
16
15
11
10
8
7
5
5
4
4
3
3
3
3
3
3
3
2
2
2
1
1
1
1
1
1
1
Wirt
Ziege
6986
5101
1927
1048
535
361
90
84
55
35
23
15
8
5
3
1
1
Klonalität
Polyklonal
Konjugat
IRDye680LT
2225
1809
1765
1261
763
687
455
310
196
155
155
150
127
122
122
102
102
96
94
89
84
79
78
75
73
68
68
68
65
61
55
55
54
54
54
53
53
53
53
53
53
53
48
48
41
38
35
34
31
24
23
23
22
20
18
18
17
16
15
15
15
14
14
14
14
14
12
12
12
12
12
10
10
10
10
10
9
9
9
8
8
8
8
8
8
7
6
6
6
6
6
5
5
5
5
5
5
4
4
4
4
4
4
4
4
4
4
3
3
3
3
3
3
3
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
1
1
1
1
1
1
1
Applikation
Immunofluorescence (IF), Immunohistochemistry (IHC), Western Blotting (WB)
Optionen
Hersteller
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Hersteller Produkt- Nr.
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'Independent Validation' Siegel
Chargennummer 27397
Validierte Anwendung Western Blotting
Positivkontrolle MCF-7 cells
Negativkontrolle SK-BR-3 cells
Bewertung A strong specific band was observed in the positive control at the expected size (~85.5 kDa) that is not observed in the negative control.
Primärantikörper
  • Antigen: Beta Catenin
  • Catalog number: ABIN1043907
  • Supplier: Anmelden zum Anzeigen
  • Lot number: 27397
  • Dilution: 1:1,000
Sekundärantikörper
Kontrollen
  • MCF-7 lysates were prepared by ADS Biosystems following standard protocols and quality controlled for protein integrity on a regular basis
  • SK-BR-3 lysates were purchased from Santa Cruz Biotechnology (catalog number sc-2218)
Protokoll
  • Lysates were mixed with NuPAGE® LDS Sample Buffer (Life Technologies NP0007) and NuPAGE® Sample Reducing Agent (Life Technologies NP0004) and denatured for 5 minutes at 90ºC.
  • 40 μg of each lysate was electrophoresed on a Bolt 4-12% Bis-Tris Gel (Life Technologies BG04120BOX) and run in Bolt MOPS SDS Running Buffer (Life Technologies B0001) at 160 volts for 1 hour.
  • Odyssey Western Protein Standard (LI-COR #928-40000) was run as a molecular weight standard.
  • PVDF membrane was activated with methanol.
  • Protein samples were transferred to activated PVDF membrane in a wet Bolt Transfer Apparatus (Life Technologies B1000) at room temperature for 1 hour at 20 volts (started at 230mA, ended at 110mA).
  • The membrane was blocked in x LI-COR Odyssey WB block solution for 1 hour at room temperature.
  • The membrane was incubated with the primary antibody diluted 1:1000 in x LI-COR Odyssey WB block solution incubated 2 hours at room temperature.
  • The membrane was washed 4 x 5 minutes in 1 x PBS-T (PBS solution with 0.1% Tween 20).
  • The membrane was incubated with IRDye® 800CW Goat anti-Mouse Secondary Antibody (Red) and IRDye 680LT Goat Anti-Rabbit Secondary Antibody (Green) from LI-COR (#827-11081, Lot #C30725-01), both 1:10,000 dilutions. Incubation was performed at room temperature for 45 minutes.
  • The membrane was washed 4 x 5 minutes in 1 x PBS-T (PBS solution with 0.1% Tween 20).
  • Proteins were detected using Odyssey machine scanning with green channel for loading control and red channel for potential LPL band.
Anmerkungen - No experimental challenges noted.
Validierungsbilder
Western Blotting validation image for anti-Catenin, beta (C-Term) antibody (ABIN1043907) Figure 1: Scanned image of Beta Catenin (Green) and loading control alpha-tubulin (Re...
'Independent Validation' Siegel
Chargennummer 19538
Validierte Anwendung Western Blotting
Positivkontrolle A549 cells
Negativkontrolle MCF-7 cells
Bewertung A strong specific band was observed in the positive control at the expected size (~175 kDa) that is not observed in the negative control.
Primärantikörper
  • Antigen: Epidermal Growth Factor Receptor (EGFR)
  • Catalog number: ABIN98862
  • Supplier: Anmelden zum Anzeigen
  • Lot number: 19538
  • Dilution: 1:1,000
Sekundärantikörper
Kontrollen - Lysates were prepared by ADS Biosystems following standard protocols and quality controlled for protein integrity on a regular basis
Protokoll
  • Lysates were mixed with NuPAGE® LDS Sample Buffer (Life Technologies NP0007) and NuPAGE® Sample Reducing Agent (Life Technologies NP0004) and denatured for 5 minutes at 90ºC.
  • 40 μg of each lysate was electrophoresed on a Bolt 4-12% Bis-Tris Gel (Life Technologies BG04120BOX) and run in Bolt MOPS SDS Running Buffer (Life Technologies B0001) at 160 volts for 1 hour.
  • Odyssey Western Protein Standard (LI-COR #928-40000) was run as a molecular weight standard.
  • PVDF membrane was activated with methanol.
  • Protein samples were transferred to activated PVDF membrane in a wet Bolt Transfer Apparatus (Life Technologies B1000) at room temperature for 1 hour at 20 volts (started at 230mA, ended at 110mA).
  • The membrane was blocked in x LI-COR Odyssey WB block solution for 1 hour at room temperature.
  • The membrane was incubated with the primary antibody diluted 1:1000 in x LI-COR Odyssey WB block solution incubated 2 hours at room temperature.
  • The membrane was washed 4 x 5 minutes in 1 x PBS-T (PBS solution with 0.1% Tween 20).
  • The membrane was incubated with IRDye® 800CW Goat anti-Mouse Secondary Antibody (Red) and IRDye 680LT Goat Anti-Rabbit Secondary Antibody (Green) from LI-COR (#827-11081, Lot #C30725-01), both 1:10,000 dilutions. Incubation was performed at room temperature for 45 minutes.
  • The membrane was washed 4 x 5 minutes in 1 x PBS-T (PBS solution with 0.1% Tween 20).
  • Proteins were detected using Odyssey machine scanning with green channel for loading control and red channel for potential LPL band.
Anmerkungen - No experimental challenges noted.
Validierungsbilder
Western Blotting validation image for anti-Epidermal Growth Factor Receptor (EGFR) antibody (ABIN98862) Figure 1: Scanned image of EGFR (Green) and loading control alpha-tubulin (Red) Weste...
Immunogen Rabbit IgG
Isotyp IgG
Spezifität Rabbit IgG
Based on ELISA and flow cytometry, this antibody reacts with the heavy and light chains of rabbit IgG, and with the light chains of rabbit IgM and IgA.
Kreuzreaktivität (Details) This antibody was tested by Dot Blot and/or solid-phase adsorbed for minimal cross-reactivity with human, mouse, rat, sheep, and chicken serum proteins, but may cross-react with immunoglobulins from other species.
Reinigung Immunoaffinity chromatography
Antigen
Applikations-hinweise Western blot: 1:20,000 Recommended
Optimum dilutions will vary and should be determined empirically.

NOTE: When using PVDF membranes for Western blotting, SDS (final concentration of 0.1 – 0.2%) and Tween® 20 (final concentration of 0.1 – 0.2%) must be added during the detection incubation step to avoid non-specific background staining.

  • IRDye 680LT secondary antibodies are suitable for immunofluorescent microscopy and other fluorescent imaging applications when using instrumentation with appropriate excitation and detection capabilities.
  • IRDye 680LT is not intended for In-Cell WesternTM assays or in vivo applications.
  • These antibodies are not recommended when getting up and running on system. Once established near-infrared protocols are optimized with IRDye 680RD, IRDye 680LT can be used to optimize signals in the 700 channel. Dilution range 1:20,000 – 1:40,000.
    Note: optimization may be required with IRDye 680LT. Please see pack insert for specific detergents/protocol modifications needed.
  • Kommentare

    • IRDye 680LT secondary antibodies have been proven to provide the brightest signal for Western blot detection in the 700 nm channel and are comparable to Alexa Fluor® 680 secondary antibodies.
    • Choose IRDye 680LT secondary antibodies to get high signal and for specific uses of detection in the 700nm channel.
    • We do not recommend to start with IRDye 680LT secondaries but rather to start with one of the IRDye 680RD secondaries.

    Beschränkungen Nur für Forschungszwecke einsetzbar
    Format Lyophilized
    Rekonstitution Reconstitute with sterile, distilled water
    Konzentration 1.0 mg/mL
    Buffer PBS, pH 7.4
    Konservierungs-mittel Sodium azide
    Vorsichtsmaßnahmen This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
    Handhabung Protect from light.
    Lagerung 4 °C