Recombinant Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Mouse monoclonal [236A/E7] to FOXP3 - BSA and Azide free
- Suitable for: IHC-P, mIHC, WB
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-FOXP3 antibody [236A/E7] - BSA and Azide free
See all FOXP3 primary antibodies -
Description
Mouse monoclonal [236A/E7] to FOXP3 - BSA and Azide free -
Host species
Mouse -
Specificity
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Tested applications
Suitable for: IHC-P, mIHC, WBmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant full length protein corresponding to FOXP3.
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Positive control
- WB: Human mammary gland lysate, HEK-293 transfected with FOXP3 expression vector containing a GFP-Myc-tag, whole cell lysate. IHC-P: Human tonsil and thymus tissue. mIHC: Human breast cancer tissue.
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General notes
This product has switched from a hybridoma to recombinant production method on 8th February 2021.
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
pH: 7.4
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
236A/E7 -
Myeloma
P3-NS1/1-Ag4-1 -
Isotype
IgG1 -
Light chain type
kappa -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab96048 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 | (2) |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.
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mIHC |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 50 kDa (predicted molecular weight: 47 kDa).
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Notes |
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IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol. |
mIHC
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 50 kDa (predicted molecular weight: 47 kDa). |
Target
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Function
Probable transcription factor. Plays a critical role in the control of immune response. -
Involvement in disease
Defects in FOXP3 are the cause of immunodeficiency polyendocrinopathy, enteropathy, X-linked syndrome (IPEX) [MIM:304790]; also known as X-linked autoimmunity-immunodeficiency syndrome. IPEX is characterized by neonatal onset insulin-dependent diabetes mellitus, infections, secretory diarrhea, trombocytopenia, anemia and eczema. It is usually lethal in infancy. -
Sequence similarities
Contains 1 C2H2-type zinc finger.
Contains 1 fork-head DNA-binding domain. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 50943 Human
- Omim: 300292 Human
- SwissProt: Q9BZS1 Human
- Unigene: 247700 Human
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Alternative names
- AIID antibody
- DIETER antibody
- Forkhead box P3 antibody
see all
Images
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Multiplex immunohistochemistry - Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)This image is courtesy of ImmunoAtlas.
Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).
Merged staining of Anti-PD-L1 (ab251611; cyan; Opal™ 520), Anti-Granzyme B (ab219803; yellow; Opal™ 540), Anti-PD1 (ab251613; magenta; Opal™ 570), Anti-pan Cytokeratin (ab264485; red; Opal™ 620), Anti-EpCAM (ab225894; red; Opal™ 620), Anti-CD8 alpha (ab251596; green; Opal™ 650) and Anti-FOXP3 (ab96048; orange; Opal™ 690). EpCAM and pan-cytokeratin share the same dye and color. Dyes are pseudo-colored for better contrast of the markers.
The immunostaining was performed on a Leica Biosystems BOND® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences®).
The section was incubated in six rounds of staining; sequentially for ab251611 (1/750 dilution), ab219803 (1/250 dilution), ab251613 (1/750 dilution), ab264485 (0.5 µg/ml), ab225894 (1/1250 dilution), ab251596 (1/1500 dilution) and ab96048 (10 µg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND® Epitope Retrieval Solution 2, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. DAPI (dark blue) was used as a nuclear counter stain.
Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra 3 Imaging System (Akoya Biosciences®).This data was developed using ab20034, the same antibody clone in a different buffer formulation.
This data is courtesy of ImmunoAtlas and it can be found here.
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Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048) at 1/1000 dilution + Human mammary gland lysate at 20 µg
Predicted band size: 47 kDaThis data was developed using ab20034, the same antibody clone in a different buffer formulation.
-
Multiplex immunohistochemistry - Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)This image is courtesy of ImmunoAtlas.
Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).
Merged staining of Anti-PD-L1 (ab251611; cyan; Opal™ 520), Anti-Granzyme B (ab219803; yellow; Opal™ 540), Anti-PD1 (ab251613; magenta; Opal™ 570), Anti-pan Cytokeratin (ab264485; red; Opal™ 620), Anti-EpCAM (ab225894; red; Opal™ 620), Anti-CD8 alpha (ab251596; green; Opal™ 650) and Anti-FOXP3 (ab96048; orange; Opal™ 690). EpCAM and pan-cytokeratin share the same dye and color. Dyes are pseudo-colored for better contrast of the markers.
The immunostaining was performed on a Leica Biosystems BOND® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences®).
The section was incubated in six rounds of staining; sequentially for ab251611 (1/750 dilution), ab219803 (1/250 dilution), ab251613 (1/750 dilution), ab264485 (0.5 µg/ml), ab225894 (1/1250 dilution), ab251596 (1/1500 dilution) and ab96048 (10 µg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND® Epitope Retrieval Solution 2, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. DAPI (dark blue) was used as a nuclear counter stain.
Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra 3 Imaging System (Akoya Biosciences®).This data was developed using ab20034, the same antibody clone in a different buffer formulation.
This data is courtesy of ImmunoAtlas and it can be found here.
-
Multiplex immunohistochemistry - Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)This image is courtesy of ImmunoAtlas.
Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).
Merged staining of Anti-PD-L1 (ab251611; cyan; Opal™ 520), Anti-Granzyme B (ab219803; yellow; Opal™ 540), Anti-PD1 (ab251613; magenta; Opal™ 570), Anti-pan Cytokeratin (ab264485; red; Opal™ 620), Anti-EpCAM (ab225894; red; Opal™ 620), Anti-CD8 alpha (ab251596; green; Opal™ 650) and Anti-FOXP3 (ab96048; orange; Opal™ 690). EpCAM and pan-cytokeratin share the same dye and color. Dyes are pseudo-colored for better contrast of the markers.
The immunostaining was performed on a Leica Biosystems BOND® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences®).
The section was incubated in six rounds of staining; sequentially for ab251611 (1/750 dilution), ab219803 (1/250 dilution), ab251613 (1/750 dilution), ab264485 (0.5 µg/ml), ab225894 (1/1250 dilution), ab251596 (1/1500 dilution) and ab96048 (10 µg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND® Epitope Retrieval Solution 2, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. DAPI (dark blue) was used as a nuclear counter stain.
Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra 3 Imaging System (Akoya Biosciences®).This data was developed using ab20034, the same antibody clone in a different buffer formulation.
This data is courtesy of ImmunoAtlas and it can be found here.
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All lanes : Anti-FOXP3 antibody [236A/E7] (ab20034) at 5 µg/ml
Lane 1 : HEK293T cell lysate
Lane 2 : HEk293T cell lysate overexpressing Human FOXP3
Lane 3 : Human tonsil tissue lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 47 kDaThis data was developed using ab20034, the same antibody clone in a different buffer formulation.
ab20034 detects Human FOXP3 protein at ~50 kDa in HEK293T cells overexpressing the protein. It also detects FOXP3 in Human tonsil tissue lysate, however this band is significantly weaker in endogenous conditions. Upon higher exposure, weak bands can also be observed in HEK293T cell lysate.
This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour before being incubated with Anti-FOXP3 antibody [236A/E7] (ab20034; 5 microgram per mL) overnight at 4°C. Antibody binding was detected using infrared labelled goat anti-mouse (green; 1:10000) for 1 hour at room temperature before imaging.
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Multiplex immunohistochemistry - Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)This image is courtesy of ImmunoAtlas.
Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).
Merged staining of Anti-PD-L1 (ab251611; cyan; Opal™ 520), Anti-Granzyme B (ab219803; yellow; Opal™ 540), Anti-PD1 (ab251613; magenta; Opal™ 570), Anti-pan Cytokeratin (ab264485; red; Opal™ 620), Anti-EpCAM (ab225894; red; Opal™ 620), Anti-CD8 alpha (ab251596; green; Opal™ 650) and Anti-FOXP3 (ab96048; orange; Opal™ 690). EpCAM and pan-cytokeratin share the same dye and color. Dyes are pseudo-colored for better contrast of the markers.
The immunostaining was performed on a Leica Biosystems BOND® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences®).
The section was incubated in six rounds of staining; sequentially for ab251611 (1/750 dilution), ab219803 (1/250 dilution), ab251613 (1/750 dilution), ab264485 (0.5 µg/ml), ab225894 (1/1250 dilution), ab251596 (1/1500 dilution) and ab96048 (10 µg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND® Epitope Retrieval Solution 2, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. DAPI (dark blue) was used as a nuclear counter stain.
Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra 3 Imaging System (Akoya Biosciences®).This data was developed using ab20034, the same antibody clone in a different buffer formulation.
This data is courtesy of ImmunoAtlas and it can be found here.
-
Multiplex immunohistochemistry - Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)This image is courtesy of ImmunoAtlas.
Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).
Merged staining of Anti-PD-L1 (ab251611; cyan; Opal™ 520), Anti-Granzyme B (ab219803; yellow; Opal™ 540), Anti-PD1 (ab251613; magenta; Opal™ 570), Anti-pan Cytokeratin (ab264485; red; Opal™ 620), Anti-EpCAM (ab225894; red; Opal™ 620), Anti-CD8 alpha (ab251596; green; Opal™ 650) and Anti-FOXP3 (ab96048; orange; Opal™ 690). EpCAM and pan-cytokeratin share the same dye and color. Dyes are pseudo-colored for better contrast of the markers.
The immunostaining was performed on a Leica Biosystems BOND® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences®).
The section was incubated in six rounds of staining; sequentially for ab251611 (1/750 dilution), ab219803 (1/250 dilution), ab251613 (1/750 dilution), ab264485 (0.5 µg/ml), ab225894 (1/1250 dilution), ab251596 (1/1500 dilution) and ab96048 (10 µg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND® Epitope Retrieval Solution 2, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. DAPI (dark blue) was used as a nuclear counter stain.
Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra 3 Imaging System (Akoya Biosciences®).This data was developed using ab20034, the same antibody clone in a different buffer formulation.
This data is courtesy of ImmunoAtlas and it can be found here.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)
This data was developed using ab20034, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling FOXP3 with ab23004 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Counterstained with hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat AntiRabbit IgG H&L (HRP). Perform heat mediated antigen retrieval using Tris/EDTA buffer (pH 9.0).
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All lanes : Anti-FOXP3 antibody [236A/E7] (ab20034) at 1/1000 dilution
Lane 1 : HEK-293 (human embryonic kidney) transfected with an empty
vector (vector control), containing a GFP-Myc-tag, whole cell lysate
Lane 2 : HEK-293 transfected with FOXP3 (WT) expression vector containing a GFP-Myc-tag, whole cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 47 kDa
Exposure time: 1 secondThis data was developed using ab20034, the same antibody clone in a different buffer formulation.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)
This data was developed using ab20034, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human thymus tissue labeling FOXP3 with ab23004 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Counterstained with hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat AntiRabbit IgG H&L (HRP). Perform heat mediated antigen retrieval using Tris/EDTA buffer (pH 9.0).
Protocols
Datasheets and documents
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Datasheet download
References (3)
ab96048 has been referenced in 3 publications.
- Krijgsman D et al. MATISSE: An analysis protocol for combining imaging mass cytometry with fluorescence microscopy to generate single-cell data. STAR Protoc 3:101034 (2022). Mass Cytometry, glucose transporter glut1 . PubMed: 34977680
- McDowell SAC et al. Neutrophil oxidative stress mediates obesity-associated vascular dysfunction and metastatic transmigration. Nat Cancer 2:545-562 (2021). PubMed: 35122017
- Sharma A et al. Anti-CTLA-4 Immunotherapy Does Not Deplete FOXP3+ Regulatory T Cells (Tregs) in Human Cancers. Clin Cancer Res 25:1233-1238 (2019). PubMed: 30054281