Recombinant Anti-GFP antibody [3H9] - BSA and Azide free (ab255886)
![Western blot - Anti-GFP antibody [3H9] - BSA and Azide free (ab255886)](https://www.abcam.com/ps/products/255/ab255886/Images/ab255886-451120-antigfp-antibody-3h9-western-blot-293t.jpg "Western blot - Anti-GFP antibody [3H9] - BSA and Azide free (ab255886)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rat monoclonal [3H9] to GFP - BSA and Azide free
- Suitable for: IHC-P, WB, ICC
- Reacts with: Species independent
Overview
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Product name
Anti-GFP antibody [3H9] - BSA and Azide free
See all GFP primary antibodies -
Description
Rat monoclonal [3H9] to GFP - BSA and Azide free -
Host species
Rat -
Specificity
This mAb recognizes eGFP, wild-type GFP, YFP and CFP.
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Tested applications
Suitable for: IHC-P, WB, ICCmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Species independent -
Immunogen
Other Immunogen Type. This information is considered to be commercially sensitive.
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Positive control
- WB: HEK-293T transfected with GFP expression vector, whole cell lysate. IHC-P: HEK-293T transfected with a GFP construct. ICC: HEK-293T cells.
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General notes
ab255886 is the carrier-free version of ab252881. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.
Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
ab255886 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.
Maxpar® is a trademark of Fluidigm Canada Inc.
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
Constituent: 100% PBS -
Carrier free
Yes -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
3H9 -
Isotype
IgG2a -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
Applications
Our Abpromise guarantee covers the use of ab255886 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| IHC-P | Use at an assay dependent concentration. | |
| WB | Use at an assay dependent concentration. Predicted molecular weight: 27 kDa. | |
| ICC | Use at an assay dependent concentration. |
Target
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Relevance
Function: Energy-transfer acceptor. Its role is to transduce the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer. Fluoresces in vivo upon receiving energy from the Ca2+ -activated photoprotein aequorin.
Subunit structure: Monomer.
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.
Biotechnological use: Green fluorescent protein has been engineered to produce a vast number of variously colored mutants, fusion proteins, and biosensors. Fluorescent proteins and its mutated allelic forms, blue, cyan and yellow have become a useful and ubiquitous tool for making chimeric proteins, where they function as a fluorescent protein tag. Typically they tolerate N- and C-terminal fusion to a broad variety of proteins. They have been expressed in most known cell types and are used as a noninvasive fluorescent marker in living cells and organisms. They enable a wide range of applications where they have functioned as a cell lineage tracer, reporter of gene expression, or as a measure of protein-protein interactions. Can also be used as a molecular thermometer, allowing accurate temperature measurements in fluids. The measurement process relies on the detection of the blinking of GFP using fluorescence correlation spectroscopy.
Sequence similarities: Belongs to the GFP family.
Biophysicochemical properties: Absorption: Abs(max)=395 nm
Exhibits a smaller absorbance peak at 470 nm. The fluorescence emission spectrum peaks at 509 nm with a shoulder at 540 nm. -
Alternative names
- GFP antibody
- Green fluorescent protein antibody
Images
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Western blot - Anti-GFP antibody [3H9] - BSA and Azide free (ab255886)All lanes : Anti-GFP antibody [3H9] (ab252881) at 1/5000 dilution
Lane 1 : HEK-293T (human embryonic kidney epithelial cell), whole cell lysate
Lane 2 : HEK-293T transfected with GFP expression vector, whole cell lysate
Lane 3 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate
Lane 4 : C6 (rat glial tumor glial cell), whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/100000 dilution
Predicted band size: 27 kDa
Observed band size: 27 kDa
Exposure time: 3 secondsThis data was developed using ab252881 the same antibody clone in a different buffer formulation.
Blocking and Diluting buffer: 5% NFDM/TBST
Exposure time: 3 seconds
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [3H9] - BSA and Azide free (ab255886)](https://www.abcam.com/ps/products/255/ab255886/Images/ab255886-451121-YR070902PSROE200wildtype293T2K.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [3H9] - BSA and Azide free (ab255886)This data was developed using ab252881 the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a GFP construct (A) labeling GFP with ab252881 at 1/2000 dilution (0.539 µg/ml) followed by a ready to use Goat Anti-rat IgG H&L (HRP polymer) (ab214882). No staining observed on HEK-293T cells (B). The section was incubated with ab252881 for 30 mins at room temperature. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is ready to use Goat Anti-rat IgG H&L (HRP polymer) (ab214882).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
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![Immunocytochemistry - Anti-GFP antibody [3H9] - BSA and Azide free (ab255886)](https://www.abcam.com/ps/products/255/ab255886/Images/ab255886-451122-293TOE487-ICC.jpg)
Immunocytochemistry - Anti-GFP antibody [3H9] - BSA and Azide free (ab255886)This data was developed using ab252881 the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cells stained for GFP (red) using ab252881 at 1/100 dilution (10.78 μg/ml), followed by ab150160 Goat Anti-Rat IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution. Confocal image showing cytoplasmic and nuclear staining in HEK-293T cells transfected with GFP only plasmid. The nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150160 Goat Anti-Rat IgG H&L (Alexa Fluor® 594) at 1/1000 dilution.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [3H9] - BSA and Azide free (ab255886)](https://www.abcam.com/ps/products/255/ab255886/Images/ab255886-451123-YR070902PSRhumantonsil2K.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [3H9] - BSA and Azide free (ab255886)This data was developed using ab252881 the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human tonsil section labeling GFP with ab252881 at 1/2000 dilution (0.539 µg/ml) followed by a ready to use Goat Anti-rat IgG H&L (HRP polymer) (ab214882). The section was incubated with ab252881 for 30 mins at room temperature. Counterstained with Hematoxylin.
Negative control: no staining on human tonsil.
Secondary antibody only control: Secondary antibody is ready to use Goat Anti-rat IgG H&L (HRP polymer) (ab214882).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [3H9] - BSA and Azide free (ab255886)](https://www.abcam.com/ps/products/255/ab255886/Images/ab255886-451124-YR070902PSRmousespleen2K.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [3H9] - BSA and Azide free (ab255886)This data was developed using ab252881 the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse spleen section labeling GFP with ab252881 at 1/2000 dilution (0.539 µg/ml) followed by a ready to use Goat Anti-rat IgG H&L (HRP polymer) (ab214882). The section was incubated with ab252881 for 30 mins at room temperature. Counterstained with Hematoxylin.
Negative control: no staining on mouse spleen.
Secondary antibody only control: Secondary antibody is ready to use Goat Anti-rat IgG H&L (HRP polymer) (ab214882).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [3H9] - BSA and Azide free (ab255886)](https://www.abcam.com/ps/products/255/ab255886/Images/ab255886-451127-YR070902PSRratliver2K.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [3H9] - BSA and Azide free (ab255886)This data was developed using ab252881 the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat liver section labeling GFP with ab252881 at 1/2000 dilution (0.539 µg/ml) followed by a ready to use Goat Anti-rat IgG H&L (HRP polymer) (ab214882). The section was incubated with ab252881 for 30 mins at room temperature. Counterstained with Hematoxylin.
Negative control: no staining on rat liver.
Secondary antibody only control: Secondary antibody is ready to use Goat Anti-rat IgG H&L (HRP polymer) (ab214882).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
References (0)
ab255886 has not yet been referenced specifically in any publications.
