TotalSeq™-Bn0001 anti-mouse CD4 Antibody, CD4, RM4-5

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Clone: RM4-5 (See other available formats)
Regulatory Status: RUO
Other Names: L3T4, T4
Isotype: Rat IgG2a, κ
Barcode Sequence: AACAAGACCCTTGAG
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Product Citations: publications

RM4-5_TotalSeq-Bn_CD4_SB_100824 — IHC staining of TotalSeq™-Bn0001 anti-mouse CD4 (clone RM4-5) on frozen mouse spleen. Following fixation with 1% PFA, the tissue was incubated with TotalSeq™-Bn001 anti-mouse CD4 (clone RM4-5) followed by incubation with Alexa Fluor® 647 Goat anti-rat IgG (Cat. No. 405416) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801) (Panel B). Images were captured using a Revvity Operetta CLS™ High Content Analysis System with a 20X objective and merged.

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100579

10 µg

369 CHF

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Description

CD4 is a 55 kD protein also known as L3T4 or T4. It is a member of the Ig superfamily, primarily expressed on most thymocytes and a subset of T cells, and weakly on macrophages and dendritic cells. It acts as a co-receptor with the TCR during T cell activation and thymic differentiation by binding MHC class II and associating with the protein tyrosine kinase lck.

Product Details

Technical Data Sheet (pdf)

Product Details

Verified Reactivity: Mouse
Antibody Type: Monoclonal
Host Species: Rat
Immunogen: BALB/c mouse thymocytes
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and EDTA
Preparation: The antibody was purified by chromatography and conjugated with TotalSeq™-Bn oligomer under optimal conditions.
Concentration: 0.5 mg/mL
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C. Do not freeze.
Application: SB - Quality tested
Recommended Usage: Each lot of this antibody is quality control tested by immunofluorescent staining in fresh frozen murine spleen tissues, and the oligomer sequence is confirmed by sequencing. TotalSeq™-Bn antibodies are compatible with the 10x Visium CytAssist Gene and Protein Expression Assay.

To maximize performance, it is strongly recommended that the reagent be titrated for each application, and that you centrifuge the antibody dilution at 14,000xg at 2 − 8°C for 10 minutes before use. Carefully pipette out the liquid avoiding the bottom of the tube when handling.

Buyer is solely responsible for determining whether Buyer has all intellectual property rights that are necessary for Buyer's intended uses of the BioLegend TotalSeq™ products. For example, for any technology platform Buyer uses with TotalSeq™, it is Buyer's sole responsibility to determine whether it has all necessary third party intellectual property rights to use that platform and TotalSeq™ with that platform.
Application Notes: The RM4-5 antibody blocks the binding of GK1.5 antibody and H129.19 antibody to CD4⁺ T cells, but not RM4-4 antibody. Additional reported applications (for the relevant formats) include: blocking of ligand binding, in vivo depletion of CD4⁺ cells¹, and immunohistochemistry of acetone-fixed frozen tissue sections^2,3,11 and paraffin-embedded sections¹¹. Clone RM4-5 is not recommended for immunohistochemistry of formalin-fixed paraffin sections. Instead, acetone frozen or zinc-fixed paraffin sections are recommended. The Ultra-LEAF™ Purified antibody (Endotoxin < 0.01 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. No. 100575 and 100576).
Additional Product Notes: TotalSeq™-Bn reagents are designed to profile protein levels following an optimized protocol in spatial transcriptomics. Compatible spatial biology devices (e.g. Imaging System, 10x Genomics Visium Spatial CytAssist Gene and Protein Expression instruments and reagents) and sequencer (e.g. Illumina analyzers) are required. TotalSeq™-B reagents are not compatible with the 10x Genomics Visium system. The complete barcode sequence may be provided upon request. Please contact technical support for more information, or visit TotalSeq™-Bn Reagents for 10x Genomics Visium CytAssist Gene and Protein Assay.

Antigen Details

Structure: Ig superfamily, 55 kD
Distribution: Majority of thymocytes, T cell subset
Function: TCR co-receptor, T cell activation
Ligand/Receptor: MHC class II molecule
Cell Type: Dendritic cells, T cells, Thymocytes, Tregs
Biology Area: Immunology
Molecular Family: CD Molecules
Antigen References: 1. Barclay A, et al. 1997. The Leukocyte Antigen FactsBook Academic Press.
2. Bierer BE, et al. 1989. Annu. Rev. Immunol. 7:579.
3. Janeway CA. 1992. Annu. Rev. Immunol. 10:645.
Gene ID: 12504 View all products for this Gene ID
UniProt: View information about CD4 on UniProt.org

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Related FAQs

I am unable to see expression of T cell markers such as CD3 and CD4 post activation.: TCR-CD3 complexes on the T-lymphocyte surface are rapidly downregulated upon activation with peptide-MHC complex, superantigen or cross-linking with anti-TCR or anti-CD3 antibodies. PMA/Ionomycin treatment has been shown to downregulate surface CD4 expression. Receptor downregulation is a common biological phenomenon and so make sure that your stimulation treatment is not causing it in your sample type.
If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?: It’s likely that other fluorophore conjugates to the same antibody clone will also be compatible with IBEX using the same sample fixation procedure. Ultimately a directly conjugated antibody’s utility in fluorescent imaging and IBEX may be specific to the sample and microscope being used in the experiment. Some antibody clone conjugates may perform better than others due to performance differences in non-specific binding, fluorophore brightness, and other biochemical properties unique to that conjugate.
Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?: Fundamentally, IBEX as a technique that works much in the same way as single antibody panels or single marker IF/IHC. If you’re already successfully using an antibody clone on a sample of interest, it is likely that clone will have utility in IBEX. It is expected some optimization and testing of different antibody fluorophore conjugates will be required to find a suitable format; however, legacy microscopy techniques like chromogenic IHC on fixed or frozen tissue is an excellent place to start looking for useful antibodies.
Are other fluorophores compatible with IBEX?: Over 18 fluorescent formats have been screened for use in IBEX, however, it is likely that other fluorophores are able to be rapidly bleached in IBEX. If a fluorophore format is already suitable for your imaging platform it can be tested for compatibility in IBEX.
The same antibody works in one tissue type but not another. What is happening?: Differences in tissue properties may impact both the ability of an antibody to bind its target specifically and impact the ability of a specific fluorophore conjugate to overcome the background fluorescent signal in a given tissue. Secondary stains, as well as testing multiple fluorescent conjugates of the same clone, may help to troubleshoot challenging targets or tissues. Using a reference control tissue may also give confidence in the specificity of your staining.
How can I be sure the staining I’m seeing in my tissue is real?: In general, best practices for validating an antibody in traditional chromogenic or fluorescent IHC are applicable to IBEX. Please reference the Nature Methods review on antibody based multiplexed imaging for resources on validating antibodies for IBEX.

Other Formats

View All CD4 Reagents Request Custom Conjugation

Description	Clone	Applications
PE/Cyanine7 anti-mouse CD4	RM4-5	FC
APC/Cyanine7 anti-mouse CD4	RM4-5	FC
APC/Fire™ 750 anti-mouse CD4	RM4-5	FC
TotalSeq™-A0001 anti-mouse CD4	RM4-5	PG
TotalSeq™-B0001 anti-mouse CD4	RM4-5	PG
TotalSeq™-C0001 anti-mouse CD4	RM4-5	PG
Ultra-LEAF™ Purified anti-mouse CD4	RM4-5	FC,CyTOF®,Depletion,Block,IHC
anti-mouse CD4 - TEST NAME CHANGE	RM4-5
anti-mouse CD4	RM4-5
APC anti-mouse CD4	RM4-5	FC
Biotin anti-mouse CD4	RM4-5	FC
FITC anti-mouse CD4	RM4-5	FC
LEAF™ Purified anti-mouse CD4	RM4-5	FC, CyTOF®, Depletion, Block, IHC
PE anti-mouse CD4	RM4-5	FC
PE/Cyanine5 anti-mouse CD4	RM4-5	FC
Purified anti-mouse CD4	RM4-5	FC,CyTOF®,IHC-F,IHC
Alexa Fluor® 647 anti-mouse CD4	RM4-5	FC,3D IHC,IHC-F
Alexa Fluor® 488 anti-mouse CD4	RM4-5	FC,IHC-F,3D IHC
Pacific Blue™ anti-mouse CD4	RM4-5	FC
Alexa Fluor® 700 anti-mouse CD4	RM4-5	FC
PerCP anti-mouse CD4	RM4-5	FC
PerCP/Cyanine5.5 anti-mouse CD4	RM4-5	FC
Brilliant Violet 421™ anti-mouse CD4	RM4-5	FC,ICC,IHC-F
Brilliant Violet 570™ anti-mouse CD4	RM4-5	FC
Brilliant Violet 605™ anti-mouse CD4	RM4-5	FC
Brilliant Violet 650™ anti-mouse CD4	RM4-5	FC
Brilliant Violet 711™ anti-mouse CD4	RM4-5	FC
Brilliant Violet 785™ anti-mouse CD4	RM4-5	FC
Brilliant Violet 510™ anti-mouse CD4	RM4-5	FC
Purified anti-mouse CD4 (Maxpar® Ready)	RM4-5	FC,CyTOF®
PE/Dazzle™ 594 anti-mouse CD4	RM4-5	FC
Spark Violet™ 423 anti-mouse CD4	RM4-5	FC
Spark Red™ 718 anti-mouse CD4 (Flexi-Fluor™)	RM4-5	FC
APC/Fire™ 810 anti-mouse CD4	RM4-5	FC
PE/Fire™ 640 anti-mouse CD4 Antibody	RM4-5	FC
Brilliant Violet 750™ anti-mouse CD4 Antibody	RM4-5	FC
Spark YG™ 581 anti-mouse CD4 (Flexi-Fluor™)	RM4-5	FC
StarBright UltraViolet 740 anti-mouse CD4	RM4-5	FC
Spark PLUS V450™ anti-mouse CD4	RM4-5	FC

Certificate of Analysis

Go To Top Version: 1 Revision Date: 10.08.2024

For Research Use Only. Not for diagnostic or therapeutic use.

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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