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

Details zu Produkt Nr. ABIN2169667, Anbieter: Anmelden zum Anzeigen
Antigen
Epitop
Heavy & Light Chain
5028
2578
1931
1069
1007
857
330
299
35
34
31
8
7
7
3
3
2
2
1
1
1
1
1
1
Reaktivität
Kaninchen
2912
2859
2405
1752
1485
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
Wirt
Ziege
6893
4912
1878
895
703
401
92
54
52
21
9
8
5
5
2
1
1
Klonalität
Polyklonal
Konjugat
IRDye680LT
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
4
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
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
Fragment Whole molecule
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
Forschungsgebiet Immunology, Secondary Antibodies
Applikationshinweise 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
    Konservierungsmittel 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