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FIGURE: 1 / 31
Mouse IgG1 Cross-Adsorbed Secondary Antibody (A-21240) in ICC/IF
Immunofluorescence analysis of Goat anti-Mouse IgG1 Cross-Adsorbed Secondary Antibody, Alexa Fluor 647 (Product # A-21240) was performed using MCF 10A cells stained with Vimentin Polyclonal Antibody (Product # PA5-27231). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, blocked with 1% BSA for 1 hour and labeled with 2 µg/mL primary antibody... View More
Please note: We are reviewing Western blot images included in the antibody testing data in our catalog, including those provided by third parties. Unless expressly labeled or annotated as “raw-unedited”, Western blot images included in the antibody testing data in our catalog may have been edited, optimized or otherwise adjusted for presentation.
Product Details
A-21240
Applications
Tested Dilution
Western Blot (WB)
1:10,000
Immunohistochemistry (Paraffin) (IHC (P))
1-10 µg/mL
Immunocytochemistry (ICC/IF)
1-10 µg/mL
Product Specifications
Species Reactivity
Mouse
Host/Isotype
Goat
/ IgG
Class
Polyclonal
Type
Secondary Antibody
Immunogen
IgG gamma 1
Conjugate
Alexa Fluor™ 647
Alexa Fluor™ 647
Alexa Fluor™ 647
Excitation/Emission Max
650/671 nm
View spectra
Form
Liquid
Concentration
2 mg/mL
Amount
500 µg
Purification
Antigen affinity chromatography
Storage buffer
PBS, pH 7.5
Contains
5mM sodium azide
Storage conditions
4°C, store in dark
Shipping conditions
Ambient (domestic); Wet ice (international)
RRID
Target
IgG1
Cross Adsorption
Prior to conjugation, antibody was solid phase adsorbed using mouse IgG2a, IgG2b and IgG3 immunosorbents to remove cross reactive antibody.
Antibody Form
Whole Antibody
Product Specific Information
To minimize cross-reactivity, these goat anti-mouse IgG1 whole secondary antibodies have been affinity purified and cross-adsorbed against mouse mouse IgM, mouse IgA, pooled human sera, purified human paraproteins, and mouse isotypes IgG2a, IgG2b, and IgG3 prior to conjugation. Cross-adsorption or pre-adsorption is a purification step to increase specificity of the antibody resulting in higher sensitivity and less background staining. The secondary antibody solution is passed through a column matrix containing immobilized serum proteins from potentially cross-reactive species. Only the nonspecific-binding secondary antibodies are captured in the column, and the highly specific secondaries flow through. The benefits of this extra step are apparent in multiplexing/multicolor-staining experiments (e.g., flow cytometry) where there is potential cross-reactivity with other primary antibodies or in tissue/cell fluorescent staining experiments where there are may be the presence of endogenous immunoglobulins.
Alexa Fluor dyes are among the most trusted fluorescent dyes available today. Invitrogen™ Alexa Fluor 647 dye is a near-infrared-fluorescent dye with excitation ideally suited to the 647 nm laser line. For stable signal generation in imaging and flow cytometry, Alexa Fluor 647 dye is pH-insensitive over a wide molar range. Probes with high fluorescence quantum yield and high photostability allow detection of low-abundance biological structures with great sensitivity. Alexa Fluor 647 dye molecules can be attached to proteins at high molar ratios without significant self-quenching, enabling brighter conjugates and more sensitive detection. The degree of labeling for each conjugate is typically 2-8 fluorophore molecules per IgG molecule; the exact degree of labeling is indicated on the certificate of analysis for each product lot.
Using conjugate solutions: Centrifuge the protein conjugate solution briefly in a microcentrifuge before use; add only the supernatant to the experiment. This step will help eliminate any protein aggregates that may have formed during storage, thereby reducing nonspecific background staining. Because staining protocols vary with application, the appropriate dilution of antibody should be determined empirically. For the fluorophore-labeled antibodies a final concentration of 1-10 µg/mL should be satisfactory for most immunohistochemistry and flow cytometry applications.
Product will be shipped at Room Temperature.
Target Information
Anti-Mouse secondary antibodies are affinity-purified antibodies with well-characterized specificity for mouse immunoglobulins and are useful in the detection, sorting or purification of its specified target. Secondary antibodies offer increased versatility enabling users to use many detection systems (e.g. HRP, AP, fluorescence). They can also provide greater sensitivity through signal amplification as multiple secondary antibodies can bind to a single primary antibody. Most commonly, secondary antibodies are generated by immunizing the host animal with a pooled population of immunoglobulins from the target species and can be further purified and modified (i.e. immunoaffinity chromatography, antibody fragmentation, label conjugation, etc.) to generate highly specific reagents.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Disclaimer
Clicking the images or links will redirect you to a website hosted by BenchSci that provides third-party scientific content. Neither the content nor the BenchSci technology and processes for selection have been evaluated by us; we are providing them as-is and without warranty of any kind, including for use or application of the Thermo Fisher Scientific products presented.
Performance Guarantee
If an Invitrogen™ antibody doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*
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