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FIGURE: 1 / 14
Rat IgG (H+L) Highly Cross-Adsorbed Secondary Antibody (A48272) in ICC/IF
Immunofluorescent analysis of tubulin in HeLa cells. The cells were fixed with 4% formaldehyde for 20 mins, washed 3x in PBS, permeabilized with 0.5% Triton X-100 in PBS for 20 minutes, washed 3x in PBS and blocked with 3% BSA in PBS for 30 minutes. Cells were incubated with Rat anti-tubulin (1:1000) and and mouse anti-ATPIF1 (1:250) (Product # A-21355) in 3% BSA in PBS overnight at 4C and washed 3x in PBS.... View More
Product Details
A48272
Applications
Tested Dilution
Western Blot (WB)
1:10,000
Immunocytochemistry (ICC/IF)
1-10 µg/mL
Product Specifications
Species Reactivity
Rat
Host/Isotype
Donkey
/ IgG
Class
Polyclonal
Type
Secondary Antibody
Immunogen
Gamma Immunoglobins Heavy and Light chains
Conjugate
Excitation/Emission Max
658/675 nm
View spectra
Form
Liquid
Concentration
2 mg/mL
Amount
1 mg
Purification
Affinity chromatography
Storage buffer
proprietary buffer, pH 6.5
Contains
0.016% Methylisothiazolone, 0.016% Bromonitrodioxane
Storage conditions
4°C, store in dark
Shipping conditions
Ambient (domestic); Wet ice (international)
RRID
Target
IgG
Cross Adsorption
against serum proteins from bovine, goat, rabbit, mouse, chicken, guinea pig, hamster, horse, sheep, and human.
Antibody Form
Whole Antibody
Product Specific Information
To minimize cross-reactivity, the donkey anti-rat IgG whole antibodies have been cross-adsorbed against serum proteins from bovine, goat, rabbit, mouse, chicken, guinea pig, hamster, horse, sheep, and human. Cross-adsorption or pre-adsorption is a purification step to increase specificity of the antibody resulting in less background staining and cross-reactivity. 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. Further passages through additional columns result in highly cross-adsorbed preparations of secondary antibody. The benefits of these extra steps are apparent in multiplexing/multicolor-staining experiments where there is potential cross-reactivity with other primary antibodies or in tissue/cell fluorescent staining experiments where there may be the presence of endogenous immunoglobulins.
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.
Specificity: This antibody binds to heavy chains on rat IgG and light chains on all rat immunoglobulins. This antibody does not bind non-immunoglobulin rat serum proteins or IgG from bovine, goat, rabbit, mouse, chicken, guinea pig, hamster, horse, sheep, or human.
Target Information
Anti-Rat secondary antibodies are affinity-purified antibodies with well-characterized specificity for rat 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.
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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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