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GFP Recombinant Rabbit Monoclonal Antibody [SP00-68]

Usd: 385

Specification

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Catalog# ET1604-25

GFP Recombinant Rabbit Monoclonal Antibody [SP00-68]

Application

  • WB

  • IHC-P

  • IF-Cell

  • IF-Tissue

  • IP

  • mIHC

Reactivity

  • Species independent

Conjugation

  • unconjugated

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

Overview

Product Name

GFP Recombinant Rabbit Monoclonal Antibody [SP00-68]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Aequorea victoria GFP aa 1-50 / 238.

Species Reactivity

Species independent

Validated Applications

WB, IHC-P, IF-Cell, IF-Tissue, IP, mIHC

Molecular Weight

Predicted band size: 27 kDa

Conjugation

unconjugated

Clone Number

SP00-68

RRID

AB_2942066

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:10,000-1:50,000

  • IF-Cell

  • 1:500-1:1,000

  • IF-Tissue

  • 1:500-1:1,000

  • IHC-P

  • 1:1,000

  • IP

  • 2-5 µg/ml.

  • mIHC

  • 1:8,000-1:10,000

Target

Function

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.

Background References

1. Yu, H. et al. 2016. AAV-Mediated Gene Transfer to Dorsal Root Ganglion. Methods in molecular biology (Clifton, N.J.). 1382: 251-61.

2. Yamaoka, M. et al. 2016. PI3K regulates endocytosis after insulin secretion by mediating signaling crosstalk between Arf6 and Rab27a. J. Cell. Sci. 129: 637-49.

Sequence Similarity

Belongs to the GFP family.

Tissue Specificity

Photocytes.

Post-translational Modification

Contains a chromophore consisting of modified amino acid residues. The chromophore is formed by autocatalytic backbone condensation between Ser-65 and Gly-67, and oxidation of Tyr-66 to didehydrotyrosine. Maturation of the chromophore requires nothing other than molecular oxygen.

UNIPROT

SwissProt: P42212 AequoreaVictoria

Synonyms

GFP antibody

Green fluorescent protein antibody

yfp antibody

Images

  • Western blot analysis of GFP on different lysates with Rabbit anti-GFP antibody (<a href="/products/ET1604-25" style="font-weight: bold;text-decoration: underline;">ET1604-25</a>) at 1/20,000 dilution.<br /><br />Lane 1: 293T cell lysate<br />Lane 2: 293T transfected with GFP cell lysate<br /><br />Lysates/proteins at 5 µg/Lane.<br /><br />Predicted band size: 27 kDa<br />Observed band size: 27 kDa<br /><br />Exposure time: 2 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1604-25" style="font-weight: bold;text-decoration: underline;">ET1604-25</a>) at 1/20,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of GFP on different lysates with Rabbit anti-GFP antibody (ET1604-25) at 1/20,000 dilution.

    Lane 1: 293T cell lysate
    Lane 2: 293T transfected with GFP cell lysate

    Lysates/proteins at 5 µg/Lane.

    Predicted band size: 27 kDa
    Observed band size: 27 kDa

    Exposure time: 2 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1604-25) at 1/20,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HeLa cells transfected with GFP labeling GFP with Rabbit anti-GFP antibody (<a href="/products/ET1604-25" style="font-weight: bold;text-decoration: underline;">ET1604-25</a>, red) at 1/500 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-GFP antibody (<a href="/products/ET1604-25" style="font-weight: bold;text-decoration: underline;">ET1604-25</a>, red) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</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 transfected with GFP labeling GFP with Rabbit anti-GFP antibody (ET1604-25, red) at 1/500 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-GFP antibody (ET1604-25, red) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) 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.

  • GFP tag was immunoprecipitated in 5µg GFP Tag fusion protein lysate with <a href="/products/ET1604-25" style="font-weight: bold;text-decoration: underline;">ET1604-25</a> at 2 µg/20 µl agarose. Western blot was performed from the immunoprecipitate using <a href="/products/M1004-8" style="font-weight: bold;text-decoration: underline;">M1004-8</a> at 1/1000 dilution. Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1:20,000 dilution was used for 60 mins at room temperature.<br /><br />Lane 1: GFP Tag fusion protein lysate (input).<br />Lane 2: Rabbit IgG instead of <a href="/products/ET1604-25" style="font-weight: bold;text-decoration: underline;">ET1604-25</a> in GFP Tag fusion protein lysate.<br />Lane 3: <a href="/products/ET1602-7" style="font-weight: bold;text-decoration: underline;">ET1602-7</a> IP in GFP Tag fusion protein lysate.<br />Lane 4: <a href="/products/ET1604-25" style="font-weight: bold;text-decoration: underline;">ET1604-25</a> IP in GFP Tag fusion protein lysate.<br />Lane 5: <a href="/products/ET1604-26" style="font-weight: bold;text-decoration: underline;">ET1604-26</a> IP in GFP Tag fusion protein lysate.<br />Lane 6: <a href="/products/ET1607-31" style="font-weight: bold;text-decoration: underline;">ET1607-31</a> IP in GFP Tag fusion protein lysate.<br />Lane 7: <a href="/products/R1312-2" style="font-weight: bold;text-decoration: underline;">R1312-2</a> IP in GFP Tag fusion protein lysate.<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST

    GFP tag was immunoprecipitated in 5µg GFP Tag fusion protein lysate with ET1604-25 at 2 µg/20 µl agarose. Western blot was performed from the immunoprecipitate using M1004-8 at 1/1000 dilution. Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:20,000 dilution was used for 60 mins at room temperature.

    Lane 1: GFP Tag fusion protein lysate (input).
    Lane 2: Rabbit IgG instead of ET1604-25 in GFP Tag fusion protein lysate.
    Lane 3: ET1602-7 IP in GFP Tag fusion protein lysate.
    Lane 4: ET1604-25 IP in GFP Tag fusion protein lysate.
    Lane 5: ET1604-26 IP in GFP Tag fusion protein lysate.
    Lane 6: ET1607-31 IP in GFP Tag fusion protein lysate.
    Lane 7: R1312-2 IP in GFP Tag fusion protein lysate.

    Blocking/Dilution buffer: 5% NFDM/TBST

  • Immunohistochemical analysis of paraffin-embedded mouse brain tissue transfected with GFP-tagged CX3CR1 with Rabbit anti-GFP antibody (<a href="/products/ET1604-25" style="font-weight: bold;text-decoration: underline;">ET1604-25</a>) at 1/1,000 dilution.<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 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1604-25" style="font-weight: bold;text-decoration: underline;">ET1604-25</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse brain tissue transfected with GFP-tagged CX3CR1 with Rabbit anti-GFP antibody (ET1604-25) at 1/1,000 dilution.

    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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1604-25) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse brain tissue transfected with GFP-tagged CX3CR1 with Rabbit anti-GFP antibody (<a href="/products/ET1604-25" style="font-weight: bold;text-decoration: underline;">ET1604-25</a>) at 1/1,000 dilution.<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 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1604-25" style="font-weight: bold;text-decoration: underline;">ET1604-25</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse brain tissue transfected with GFP-tagged CX3CR1 with Rabbit anti-GFP antibody (ET1604-25) at 1/1,000 dilution.

    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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1604-25) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

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Citation