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Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 647

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Immunofluorescent analysis of ZO-1 in A549 cells. The cells were fixed with 4% formaldehyde for 15 mins, permeabilized with 0.25% Triton X-100 in PBS for 10 mins, and blocked with 3% BSA in PBS for 30 mins at RT. Cells were stained with a ZO-1 rabbit antibody (Product # 40-2300) at a dilution of 1:200 in 3% BSA in PBS for 1 hr at RT, and then incubated with Invitrogen Alexa Fluor Plus 647 goat anti-rabbit IgG secondary antibody (Product # A32733) at a dilution of 1:1000 for 1 hr at RT. Nuclei were stained with Hoechst 33342 (Product # H3570). The image contains overlay of ZO-1 (far red) and nuclei (blue). Images were taken on a Zeiss LSM 710 confocal microscope at 40X magnification.

Catalog No - A32733
HSN - 38229090
₹ 36,649.00 ₹ 36,649.00  ​

₹ 36,649.00 / Units
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​Brand: Invitrogen

               


                  Product Details

Product Specific Information

To minimize cross-reactivity, the goat anti-rabbit IgG whole antibodies have been pre cross-adsorbed against bovine IgG, goat IgG, mouse IgG, rat IgG, and human IgG. 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.

Target Information

Anti-Rabbit secondary antibodies are affinity-purified antibodies with well-characterized specificity for rabbit 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.

               Specifications


Species Reactivity

Rabbit

Host/Isotype

Goat / IgG

Class

Polyclonal

Type

Secondary Antibody

Immunogen

Gamma Immunoglobins Heavy and Light chains

Conjugate

Alexa Fluor™ Plus 647

Excitation/Emission Max

658/675 nm

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

AB_2633282

Target

IgG

Cross Adsorption

Against bovine IgG, goat IgG, mouse IgG, rat IgG and human IgG

Antibody Form

Whole Antibody

Applications

Tested Dilution

Western Blot (WB)

0.05-0.2 µg/mL

Immunohistochemistry (Paraffin) (IHC (P)

Assay-dependent

Immunohistochemistry (Frozen) (IHC (F))

Assay-dependent

Immunocytochemistry (ICC/IF)

1-10 µg/mL



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