Beads抗体-Anti-GFP-Tag Antibody (Agarose conjugated)

Datasheet

Background收起

The green fluorescent protein (GFP) was originally identified as a protein involved in the bioluminescence of the jellyfish Aequorea victoria. GFP cDNA produces a fluorescent product when expressed in prokaryotic cells, without the need for exogenous substrates or cofactors, making GFP a useful tool for monitoring gene expression and protein localization in vivo. Several GFP mutants have been developed, including EGFP, which fluoresce more intensely than the wildtype GFP and have shifted excitation maxima, making them useful for FACS and fluorescence microscopy as well as double-labeling applications. GFP is widely used in expression vectors as a fusion protein tag, allowing expression and monitoring of heterologous proteins fused to GFP.

Immunogen收起

This Abmart monoclonal antibody is produced by immunizing animals with full length recombinant GFP.

Isotype收起

Mouse IgG1

Specificity收起

GFP-Tag (7G9) Mouse mAb detects GFP and GFP fusion proteins.

ReActivity收起

All

Application Image收起

  • WB
  • IP
  • 企业微信截图_15760555407104.png

  • The work can be performed in 1.5 ml micro-centrifuge tubes or in spin columns.

    1. Thoroughly resuspend the Anti-HA Agarose by inverting the tube or vial several times.

    2. Add 20-50 μl 50% slurry of Anti-HA Agarose into cell lysate using a widebore pipette tip.

    Note: The lysate should be fresh, and for a well expressed tagged protein, 200 μl lysate (200-500 μg total protein) usually yields a good IP result.

    3. Incubate with gentle mixing for 2 h to overnight at 4 .

    4. Wash the beads with 1 ml TBS buffer or lysis buffer, such as RIPA (50 mM Tris HCl, pH 7.4, 150 mM NaCl, 1 mM EDTA, 1% NP-40, 0.5% sodium deoxycholate), centrifuge for 3 min at 2,000 g, and discard the supernatant. Wash 3 times, avoid losing beads during washes.

    5. Elution of the HA tagged protein.

    Option 1. Elution with elution buffer.

    Add 30-50 μl elution buffer to the beads, gently tap the tube to mix well, immediately centrifuge for 3 min, transfer the supernatant very carefully to a fresh tube (Avoid transferring any beads).

    Note: Neutralize the eluant immediately by add 1 l of 1.5 M Tris, pH 9.0 per 20 μl Elution buffer.

    Option 2. Elution with HA peptide

    Add 30-50 μl HA peptide solution (100 μg/ml HA peptide in TBS buffer), gently tap the tube to mix well, incubate for 10 min, centrifuge for 3 min, and transfer the supernant to a fresh tube. TBS buffer: 50 mM Tris HCl, 150 mM NaCl, pH 7.4.

    Option 3. Elution with SDS loading buffer

    Add 30 μl 2 SDS loading buffer, gently tap the tube to mix well, boil at 100 °C for 5 min, centrifuge for 3 min, transfer the supernatant to a fresh tube.

    Note: in this case, the supernatant contains not only the binding proteins, but also IgG (heavy and light chains).

    6. Prepare SDS-PAGE gel for western blotting or proceed to other assays.

Storage Condition收起

Theproductissuppliedasa50%slurryinstoragebuffer(1PBS,pH7.4,containing0.1%NaN).

Storetheproductat4°Canddonotfreeze.

RRID收起

AB_2936243

References

M20014

货号 规格 价格(¥)
M20014S 500μl 2,650
M20014M 1000μl 3,300
M20014L 5000μl 10,800

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Note:  Application as IHC, only suitable for histochemical staining or fluorescence staining of paraffin-embedded sections. Application as ICC/IF, suitable for histochemical or fluorescent staining of frozen sections, as well as chemical and fluorescent staining at the cellular level.

注意:抗体应用为IHC的,抗体只适合于石蜡切片的组化染色或者荧光染色。

抗体应用为IF/ICC的,抗体适合于冰冻切片的组化染色或者荧光染色,以及细胞水平的化学染色和荧光染色。

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应聘职位:hr@ab-mart.com

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销售电话:13162017139(微信同号)

技术支持:15618194176(微信同号)

总机:021-34695901

南方经销商负责:手机13122837132(微信同号)          
北方及西南经销商负责:手机13122150513(微信同号)

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