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iFluor™ 488 Conjugated Goat anti-rabbit IgG polyclonal Antibody

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Catalog# HA1121

iFluor™ 488 Conjugated Goat anti-rabbit IgG polyclonal Antibody

Application

  • IF-Cell

  • IF-Tissue

  • FC

  • IHC-Fr

  • WB

Reactivity

  • Rabbit

Conjugation

  • iFluor™ 488

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

製品概要

製品名

iFluor™ 488 Conjugated Goat anti-rabbit IgG polyclonal Antibody

抗体のタイプ

Goat Polyclonal Antibody

免疫原

Rabbit IgG (H+L).

製品の特異性

To minimize cross-reactivity, the iFluor™ 488 conjugated goat anti-rabbit IgG (H+L) antibodies have been highly cross-adsorbed against human IgG and Mouse IgG.

交差反応性（対応種属）

Rabbit

検証済みアプリケーション

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

コンジュゲーション

iFluor™ 488

製品の特徴

形態

Liquid

濃度

保存方法

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

保存バッファー

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

アイソタイプ

IgG

精製方法

Immunogen affinity purified.

推奨希釈倍率

  • IF-Cell

  • 1:500-1:1,000

  • IF-Tissue

  • 1:500-1:1,000

  • FC

  • 1:500-1:1,000

  • IHC-Fr

  • 1:500-1:1,000

  • WB

  • 1:5,000

ターゲット

機能

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. Although FITC is still the most popular fluorescent labeling dye for preparing green fluorescent bioconjugates, there are certain limitations with FITC, such as severe photobleaching for microscope imaging and pH-sensitive fluorescence. Protein conjugates prepared with iFluor™ 488 dyes are far superior compared to conjugates of fluorescein derivatives such as FITC. iFluor™ 488 conjugates are significantly brighter than fluorescein conjugates and are much more photostable. Additionally, the fluorescence of iFluor™ 488 is not affected by pH (4-10). This pH insensitivity is a major improvement over fluorescein, which emits its maximum fluorescence only at pH above 9. iFluor™ 488 SE dye is reasonably stable and shows good reactivity and selectivity with protein amino groups.

背景・参考文献

暂无

画像

  • Application: Immunofluorescence (IHC-Fr)<br /><br />Species: Mouse<br />Tissue: Cerebellum<br />Sample: Frozen section<br /><br />Antigen retrieval: Recommend. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven.<br /><br />Wash buffer: 1× TBST<br />Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.<br /><br />Primary antibody: <a href="/products/HA723738" style="font-weight: bold;text-decoration: underline;">HA723738</a>, 1/5,000, overnight at 4℃.<br />Secondary antibody: Goat anti-Rabbit IgG (iFluor&trade; 488), <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>, 1/1,000, 1.5 hours at room temperature.

    Application: Immunofluorescence (IHC-Fr)

    Species: Mouse
    Tissue: Cerebellum
    Sample: Frozen section

    Antigen retrieval: Recommend. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven.

    Wash buffer: 1× TBST
    Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.

    Primary antibody: HA723738, 1/5,000, overnight at 4℃.
    Secondary antibody: Goat anti-Rabbit IgG (iFluor™ 488), HA1121, 1/1,000, 1.5 hours at room temperature.

  • Application: Immunofluorescence (IHC-Fr)<br /><br />Species: Rat<br />Tissue: Cerebellum<br />Sample: Frozen section<br /><br />Antigen retrieval: Recommend. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven.<br /><br />Wash buffer: 1× TBST<br />Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.<br /><br />Primary antibody: <a href="/products/HA723738" style="font-weight: bold;text-decoration: underline;">HA723738</a>, 1/5,000, overnight at 4℃.<br />Secondary antibody: Goat anti-Rabbit IgG (iFluor&trade; 488), <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>, 1/1,000, 1.5 hours at room temperature.

    Application: Immunofluorescence (IHC-Fr)

    Species: Rat
    Tissue: Cerebellum
    Sample: Frozen section

    Antigen retrieval: Recommend. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven.

    Wash buffer: 1× TBST
    Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.

    Primary antibody: HA723738, 1/5,000, overnight at 4℃.
    Secondary antibody: Goat anti-Rabbit IgG (iFluor™ 488), HA1121, 1/1,000, 1.5 hours at room temperature.

  • Application: Immunofluorescence (IF-tissue)<br /><br />Species: Rat<br />Tissue: Cerebellum<br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, sodium citrate buffer (pH 6.0), 2 minutes at high pressure.<br /><br />Wash buffer: 1× TBST<br />Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.<br />Primary antibody: Antibody A: 1/1,000 (NeuN , <a href="/products/ET1602-12" style="font-weight: bold;text-decoration: underline;">ET1602-12</a>, rabbit, Green), Antibody B: 1/1,000 (GFAP, <a href="/products/EM140707" style="font-weight: bold;text-decoration: underline;">EM140707</a>, mouse, Red), overnight at 4℃. <br />Secondary antibody: Goat anti-Rabbit IgG (iFluor&trade; 488), <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>, 1/1,000; Goat anti-Mouse IgG (iFluor&trade; 594), <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>, 1/1,000, 1.5 hours at room temperature.

    Application: Immunofluorescence (IF-tissue)

    Species: Rat
    Tissue: Cerebellum
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, sodium citrate buffer (pH 6.0), 2 minutes at high pressure.

    Wash buffer: 1× TBST
    Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
    Primary antibody: Antibody A: 1/1,000 (NeuN , ET1602-12, rabbit, Green), Antibody B: 1/1,000 (GFAP, EM140707, mouse, Red), overnight at 4℃.
    Secondary antibody: Goat anti-Rabbit IgG (iFluor™ 488), HA1121, 1/1,000; Goat anti-Mouse IgG (iFluor™ 594), HA1126, 1/1,000, 1.5 hours at room temperature.

  • Application: Immunocytochemistry (IF-cell)<br /><br />Species: Human<br />Sample: HeLa (Human cervix adenocarcinoma epithelial cell)<br /><br />Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.<br />Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.<br />Antibody dilution buffer: 1% BSA in PBST.<br /><br />Primary antibody: beta Actin (<a href="/products/HA722023" style="font-weight: bold;text-decoration: underline;">HA722023</a>), 1/250, overnight at 4℃.<br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), , 1/1,000, 45 minutes at room temperature.<br />Counterstain: Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

    Application: Immunocytochemistry (IF-cell)

    Species: Human
    Sample: HeLa (Human cervix adenocarcinoma epithelial cell)

    Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
    Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
    Antibody dilution buffer: 1% BSA in PBST.

    Primary antibody: beta Actin (HA722023), 1/250, overnight at 4℃.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), , 1/1,000, 45 minutes at room temperature.
    Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

  • Application: Immunocytochemistry (IF-cell)<br /><br />Species: Human<br />Sample: NIH/3T3 (Mouse fibroblast)<br /><br />Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.<br />Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.<br />Antibody dilution buffer: 1% BSA in PBST.<br /><br />Primary antibody: beta Actin (<a href="/products/HA722023" style="font-weight: bold;text-decoration: underline;">HA722023</a>), 1/100, overnight at 4℃.<br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 1/1,000, 45 minutes at room temperature.<br />Counterstain: Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

    Application: Immunocytochemistry (IF-cell)

    Species: Human
    Sample: NIH/3T3 (Mouse fibroblast)

    Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
    Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
    Antibody dilution buffer: 1% BSA in PBST.

    Primary antibody: beta Actin (HA722023), 1/100, overnight at 4℃.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1/1,000, 45 minutes at room temperature.
    Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

  • Application: Immunocytochemistry (IF-cell)<br /><br />Species: Human<br />Sample: L6 (Rat skeletal muscle myoblast)<br /><br />Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.<br />Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.<br />Antibody dilution buffer: 1% BSA in PBST.<br /><br />Primary antibody: beta Actin (<a href="/products/HA722023" style="font-weight: bold;text-decoration: underline;">HA722023</a>), 1/100, overnight at 4℃.<br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), , 1/1,000, 45 minutes at room temperature.<br />Counterstain: Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

    Application: Immunocytochemistry (IF-cell)

    Species: Human
    Sample: L6 (Rat skeletal muscle myoblast)

    Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
    Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
    Antibody dilution buffer: 1% BSA in PBST.

    Primary antibody: beta Actin (HA722023), 1/100, overnight at 4℃.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), , 1/1,000, 45 minutes at room temperature.
    Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

  • Application: Flow Cytometry<br /><br />Species: Human<br />Sample: MOLT-4 (Human acute T-Cell leukemia cell)<br /><br />Blocking: 1% BSA + 10% normal goat serum + human FcR (Miltenyi Biotec), 15 minutes at room temperature. <br />Antibody dilution buffer: 1x PBS.<br /><br />Primary antibody: CD3 (<a href="/products/HA724302" style="font-weight: bold;text-decoration: underline;">HA724302</a>, 1 μg/mL, Red) compared with Rabbit IgG Isotype Control (<a href="/products/HA722127" style="font-weight: bold;text-decoration: underline;">HA722127</a>, Green), 15 minutes at room temperature.<br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488), <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>, 1/1,000, 15 minutes at room temperature.

    Application: Flow Cytometry

    Species: Human
    Sample: MOLT-4 (Human acute T-Cell leukemia cell)

    Blocking: 1% BSA + 10% normal goat serum + human FcR (Miltenyi Biotec), 15 minutes at room temperature.
    Antibody dilution buffer: 1x PBS.

    Primary antibody: CD3 (HA724302, 1 μg/mL, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488), HA1121, 1/1,000, 15 minutes at room temperature.

  • Application: Flow Cytometry (Intra)<br /><br />Species: Human<br />Sample: HeLa (Human cervix adenocarcinoma epithelial cell)<br /><br />Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.<br />Permeabilization: 0.1% Tween-20, 15 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.<br />Antibody dilution buffer: 1x PBS.<br /><br />Primary antibody: beta actin (<a href="/products/HA722023" style="font-weight: bold;text-decoration: underline;">HA722023</a>, 1 μg/mL, Red) compared with Rabbit IgG Isotype Control (<a href="/products/HA722127" style="font-weight: bold;text-decoration: underline;">HA722127</a>, Green), 15 minutes at room temperature.<br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 1/1,000, 15 minutes at room temperature.

    Application: Flow Cytometry (Intra)

    Species: Human
    Sample: HeLa (Human cervix adenocarcinoma epithelial cell)

    Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
    Permeabilization: 0.1% Tween-20, 15 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
    Antibody dilution buffer: 1x PBS.

    Primary antibody: beta actin (HA722023, 1 μg/mL, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1/1,000, 15 minutes at room temperature.

  • Western blot analysis of Goat Anti-Rabbit IgG - iFluor&trade; 488 Secondary Antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/5,000 dilution.<br /><br />Lane 1: MCF7 (Human breast cancer cell) cell lysate (20 µg/Lane)<br />Lane 2: Jurkat (Human T-lymphoblastic cells) cell lysate (20 µg/Lane)<br />Lane 3: Mouse testis tissue lysate (40 µg/Lane)<br />Lane 4: Rat heart tissue lysate (40 µg/Lane)<br /><br />Exposure time: 2 seconds; ECL: K1801<br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: Hsp90 beta (<a href="/products/ET1605-56" style="font-weight: bold;text-decoration: underline;">ET1605-56</a>), 1/20,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), 2 hours at room temperature<br />Secondary antibody: Goat anti-Rabbit IgG (iFluor&trade; 488), <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>, 1/5,000 in 5% NFDM/TBST, 1 hour at room temperature

    Western blot analysis of Goat Anti-Rabbit IgG - iFluor™ 488 Secondary Antibody (HA1121) at 1/5,000 dilution.

    Lane 1: MCF7 (Human breast cancer cell) cell lysate (20 µg/Lane)
    Lane 2: Jurkat (Human T-lymphoblastic cells) cell lysate (20 µg/Lane)
    Lane 3: Mouse testis tissue lysate (40 µg/Lane)
    Lane 4: Rat heart tissue lysate (40 µg/Lane)

    Exposure time: 2 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: Hsp90 beta (ET1605-56), 1/20,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature
    Secondary antibody: Goat anti-Rabbit IgG (iFluor™ 488), HA1121, 1/5,000 in 5% NFDM/TBST, 1 hour at room temperature

文献・引用論文

ご注意: 本製品はすべて「研究用試薬」です。人や動物の診断・治療目的、または臨床診断には使用できません。（FOR RESEARCH USE ONLY）

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