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    products/primary-antibodies/foxp3-antibody-236ae7-bsa-and-azide-free-ab96048.pdf

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Recombinant

Recombinant Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)

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Multiplex immunohistochemistry - Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)
  • Western blot - Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)
  • Multiplex immunohistochemistry - Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)
  • Multiplex immunohistochemistry - Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)
  • Western blot - Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)
  • Multiplex immunohistochemistry - Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)
  • Multiplex immunohistochemistry - Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)
  • Western blot - Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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

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Overview

  • 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

    The epitope recognized by FOXP3 antibody [236A/E7] (ab20034) is between amino acids 105-236. FOXP3 antibody [236A/E7] (ab20034) is expected to detect full length FOXP3 as well as both cleaved forms.

  • Tested applications

    Suitable for: IHC-P, mIHC, WBmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant full length protein corresponding to FOXP3.

  • 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.
  • 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
    For more information see here.

Properties

  • 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...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    236A/E7
  • Myeloma

    P3-NS1/1-Ag4-1
  • Isotype

    IgG1
  • Light chain type

    kappa
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Cell Division
    • Chromatid Cohesion
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Domain Families
    • Forkhead Box
    • FOXP
    • Immunology
    • Adaptive Immunity
    • Regulatory T Cells

Associated products

  • Alternative Versions

    • Anti-FOXP3 antibody [236A/E7] (ab20034)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
    • Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)
  • Recombinant Protein

    • Recombinant Human FOXP3 protein (His tag) (ab226445)

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 (1)
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).
Notes
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

  • 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.
  • Target information above from: UniProt accession Q9BZS1 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 50943 Human
    • Omim: 300292 Human
    • SwissProt: Q9BZS1 Human
    • Unigene: 247700 Human
    • Alternative names

      • AIID antibody
      • DIETER antibody
      • Forkhead box P3 antibody
      • Forkhead box protein P3 antibody
      • FOXP3 antibody
      • FOXP3_HUMAN antibody
      • FOXP3delta7 antibody
      • Immune dysregulation polyendocrinopathy enteropathy X linked antibody
      • Immunodeficiency polyendocrinopathy enteropathy X linked antibody
      • IPEX antibody
      • JM2 antibody
      • MGC141961 antibody
      • MGC141963 antibody
      • OTTHUMP00000025832 antibody
      • OTTHUMP00000025833 antibody
      • OTTHUMP00000226737 antibody
      • PIDX antibody
      • Scurfin antibody
      • XPID antibody
      see all

    Images

    • Multiplex immunohistochemistry - Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)
      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), Anti-Granzyme B (ab219803), Anti-PD1 (ab251613), Anti-pan Cytokeratin (ab264485), Anti-EpCAM (ab225894), Anti-CD8 alpha (ab251596) and Anti-FOXP3 (ab96048).

      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.

    • Western blot - Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)
      Western blot - Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)
      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 kDa



      This 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)
      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), Anti-Granzyme B (ab219803), Anti-PD1 (ab251613), Anti-pan Cytokeratin (ab264485), Anti-EpCAM (ab225894), Anti-CD8 alpha (ab251596) and Anti-FOXP3 (ab96048).

      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)
      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), Anti-Granzyme B (ab219803), Anti-PD1 (ab251613), Anti-pan Cytokeratin (ab264485), Anti-EpCAM (ab225894), Anti-CD8 alpha (ab251596) and Anti-FOXP3 (ab96048).

      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.

    • Western blot - Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)
      Western blot - Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)
      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 kDa



      This 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.

    • Multiplex immunohistochemistry - Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)
      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), Anti-Granzyme B (ab219803), Anti-PD1 (ab251613), Anti-pan Cytokeratin (ab264485), Anti-EpCAM (ab225894), Anti-CD8 alpha (ab251596) and Anti-FOXP3 (ab96048).

      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)
      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), Anti-Granzyme B (ab219803), Anti-PD1 (ab251613), Anti-pan Cytokeratin (ab264485), Anti-EpCAM (ab225894), Anti-CD8 alpha (ab251596) and Anti-FOXP3 (ab96048).

      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)
      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).

    • Western blot - Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)
      Western blot - Anti-FOXP3 antibody [236A/E7] - BSA and Azide free (ab96048)
      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 second


      This 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)
      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

    • Flow cytometry protocols
    • Immunohistochemistry protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (2)

    Publishing research using ab96048? Please let us know so that we can cite the reference in this datasheet.

    ab96048 has been referenced in 2 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
    • 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

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-FOXP3 antibody [236A/E7] - BSA and Azide free

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Melanoma lymph node)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris-EDTA PH 9
    Permeabilization
    Yes - Tween
    Specification
    Melanoma lymph node
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    no additional fixation
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Apr 21 2021

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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