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iFluor™ 647 Conjugated Goat anti-rabbit IgG polyclonal Antibody

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Catalog# HA1123

iFluor™ 647 Conjugated Goat anti-rabbit IgG polyclonal Antibody

Application

  • IF-Cell

  • IF-Tissue

  • IHC-Fr

  • FC

Reactivity

  • Rabbit

Conjugation

  • iFluor™ 647

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

iFluor™ 647 Conjugated Goat anti-rabbit IgG polyclonal Antibody

Antibody Type

Goat Polyclonal Antibody

Immunogen

Rabbit IgG (H+L).

Product Specificity

To minimize cross-reactivity, the iFluor™ 647 conjugated goat anti-rabbit IgG (H+L) antibodies have been highly cross-adsorbed against human IgG and Mouse IgG.

Species Reactivity

Rabbit

Validated Applications

IF-Cell, IF-Tissue, IHC-Fr, FC

Conjugation

iFluor™ 647

Product Features

Form

Liquid

Concentration

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4), 1% BSA, 40% Glycerol, 0.2% Proclean 950.

Isotype

IgG

Purification Method

Immunogen affinity purified.

Application Dilution

  • IF-Cell

  • 1:500-1:1,000

  • IF-Tissue

  • 1:500-1:1,000

  • IHC-Fr

  • 1:500-1:1,000

  • FC

  • 1:1,000

Target

Function

Whole IgG antibodies are isolated as intact molecules from antisera by immunoaffinity chromatography. They have an Fc portion and two antigen binding Fab portions joined together by disulfide bonds and therefore they are divalent. The average molecular weight is reported to be about 160 kDa. The whole IgG form of antibodies is suitable for the majority of immunodetection procedures and is the most cost effective. iFluor™ 647 dyes have fluorescence excitation and emission maxima of ~656 nm and ~670 nm respectively. iFluor™ 647 family is pH-independent from pH 3 to 11. These spectral characteristics make this new dye family an excellent alternative. iFluor™ 647 is much easier to be conjugated with RPE with much higher conjugation yield, and the resulted RPE-iFluor™ 647 tandem has better FRET efficiency. iFluor™ 647 SE is reasonably stable and shows good reactivity and selectivity with protein amino groups.

Background References

暂无

Images

  • Immunocytochemistry analysis of Hela cells labeling Vimentin with Rabbit anti-Vimentin antibody (<a href="/products/ET1610-39" style="font-weight: bold;text-decoration: underline;">ET1610-39</a>) at 1/400 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 3% BSA for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Vimentin antibody (<a href="/products/ET1610-39" style="font-weight: bold;text-decoration: underline;">ET1610-39</a>) at 1/400 dilution in 3% BSA overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 647, <a href="/products/HA1123" style="font-weight: bold;text-decoration: underline;">HA1123</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of Hela cells labeling Vimentin with Rabbit anti-Vimentin antibody (ET1610-39) at 1/400 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 3% BSA for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Vimentin antibody (ET1610-39) at 1/400 dilution in 3% BSA overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 647, HA1123) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

  • Immunofluorescence analysis of paraffin-embedded mouse brain tissue labeling Synaptophysin (<a href="/products/ET1606-56" style="font-weight: bold;text-decoration: underline;">ET1606-56</a>) and beta Ⅲ tubulin (<a href="/products/M0805-8" style="font-weight: bold;text-decoration: underline;">M0805-8</a>).<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS. And then probed with the primary antibodies Synaptophysin (<a href="/products/ET1606-56" style="font-weight: bold;text-decoration: underline;">ET1606-56</a>, red) at 1/200 dilution and beta Ⅲ tubulin (<a href="/products/M0805-8" style="font-weight: bold;text-decoration: underline;">M0805-8</a>, green) at 1/200 dilution overnight at 4 ℃, washed with PBS.<br /><br />iFluor&trade; 647 conjugate-Goat anti-Rabbit IgG (<a href="/products/HA1123" style="font-weight: bold;text-decoration: underline;">HA1123</a>) and iFluor&trade; 488 conjugate-Goat anti-Mouse IgG (<a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) were used as the secondary antibodies at 1/500 dilution. DAPI was used as nuclear counterstain.

    Immunofluorescence analysis of paraffin-embedded mouse brain tissue labeling Synaptophysin (ET1606-56) and beta Ⅲ tubulin (M0805-8).

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS. And then probed with the primary antibodies Synaptophysin (ET1606-56, red) at 1/200 dilution and beta Ⅲ tubulin (M0805-8, green) at 1/200 dilution overnight at 4 ℃, washed with PBS.

    iFluor™ 647 conjugate-Goat anti-Rabbit IgG (HA1123) and iFluor™ 488 conjugate-Goat anti-Mouse IgG (HA1125) were used as the secondary antibodies at 1/500 dilution. DAPI was used as nuclear counterstain.

  • Flow cytometric analysis of HeLa cells labeling beta Tubulin.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 647 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1123" style="font-weight: bold;text-decoration: underline;">HA1123</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HeLa cells labeling beta Tubulin.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1602-4, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 647 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1123) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Citation

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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