- GK1.5 (See other available formats)
- Regulatory Status
- Other Names
- L3T4, T4
- Rat IgG2b, κ
- Ave. Rating
- Submit a Review
- Product Citations
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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, a subset of T cells, and weakly on macrophages and dendritic cells. It acts as a coreceptor with the TCR during T cell activation and thymic differentiation by binding MHC class II and associating with the protein tyrosin kinase, lck.Product Details
- Verified Reactivity
- Antibody Type
- Host Species
- Mouse CTL clone V4
- Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
- The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 594 under optimal conditions.
- 0.5 mg/ml
- Storage & Handling
- The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
IHC-F - Quality tested
FC - Verified
SB - Reported in the literature, not verified in house
- Recommended Usage
Each lot of this antibody is quality control tested by immunohistochemistry. For immunohistochemistry on frozen tissue sections, a concentration range of 2.5-5 μg/ml is suggested. For flow cytometric staining, the suggested use of this reagent is ≤0.25 µg per million cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.
* Alexa Fluor® 594 has an excitation maximum of 590 nm, and a maximum emission of 617 nm.
Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.
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- Application Notes
Additional reported applications (for the relevant formats) include: blocking of CD4+ T cell activation1,4,11, thymocyte costimulation3, in vitro and in vivo depletion2,5-8, blocking of egg-sperm cell adhesion1,4, immunohistochemical staining of acetone-fixed frozen sections9,10, immunoprecipitation1,2, and spatial biology (IBEX)12,13. The GK1.5 antibody is able to block CD4 mediated cell adhesion and T cell activation. Binding of GK1.5 antibody to CD4 T cells can be blocked by RM4-5 antibody, but not RM4-4 antibody. For in vivo studies or highly sensitive assays, we recommend Ultra-LEAF™ purified antibody (Cat. No. 100442) with a lower endotoxin limit than standard LEAF™ purified antibodies (Endotoxin < 0.01 EU/µg).
- Additional Product Notes
Iterative Bleaching Extended multi-pleXity (IBEX) is a fluorescent imaging technique capable of highly-multiplexed spatial analysis. The method relies on cyclical bleaching of panels of fluorescent antibodies in order to image and analyze many markers over multiple cycles of staining, imaging, and, bleaching. It is a community-developed open-access method developed by the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
(PubMed link indicates BioLegend citation)
- Dialynas DP, et al. 1983. J. Immunol. 131:2445. (Block, IP)
- Dialynas DP, et al. 1983. Immunol. Rev. 74:29. (IP, Deplete)
- Wu L, et al. 1991. J. Exp. Med. 174:1617. (Costim)
- Godfrey DI, et al. 1994. J. Immunol. 152:4783. (Block)
- Gavett SH, et al. 1994. Am. J. Respir. Cell. Mol. Biol. 10:587. (Deplete)
- Schuyler M, et al. 1994. Am. J. Respir. Crit. Care Med. 149:1286. (Deplete)
- Ghobrial RR, et al. 1989. Clin. Immunol. Immunopathol. 52:486. (Deplete)
- Israelski DM, et al. 1989. J. Immunol. 142:954. (Deplete)
- Zheng B, et al. 1996. J. Exp. Med. 184:1083. (IHC)
- Frei K, et al. 1997. J. Exp. Med. 185:2177. (IHC)
- Felix NJ, et al. 2007. Nat. Immunol. 8:388. (Block)
- Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
- Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
- Product Citations
AB_2563182 (BioLegend Cat. No. 100446)
- Ig superfamily, 55 kD
Majority of thymocytes, T cell subset
- TCR co-receptor, T cell activation
- MHC class II molecule
- Cell Type
- Dendritic cells, T cells, Thymocytes, Tregs
- Biology Area
- 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
- View information about CD4 on UniProt.org
- 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.
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Compare Data Across All Formats
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.
If you need assistance with selecting the best format contact our expert technical support team.
APC anti-mouse CD4
Biotin anti-mouse CD4
FITC anti-mouse CD4
PE anti-mouse CD4
PE/Cyanine5 anti-mouse CD4
Purified anti-mouse CD4
PE/Cyanine7 anti-mouse CD4
APC/Cyanine7 anti-mouse CD4
Alexa Fluor® 647 anti-mouse CD4
Alexa Fluor® 488 anti-mouse CD4
Pacific Blue™ anti-mouse CD4
Alexa Fluor® 700 anti-mouse CD4
PerCP anti-mouse CD4
PerCP/Cyanine5.5 anti-mouse CD4
Brilliant Violet 421™ anti-mouse CD4
Ultra-LEAF™ Purified anti-mouse CD4
Alexa Fluor® 594 anti-mouse CD4
Brilliant Violet 711™ anti-mouse CD4
Brilliant Violet 510™ anti-mouse CD4
Brilliant Violet 605™ anti-mouse CD4
Brilliant Violet 785™ anti-mouse CD4
PE/Dazzle™ 594 anti-mouse CD4
APC/Fire™ 750 anti-mouse CD4
GoInVivo™ Purified anti-mouse CD4
Brilliant Violet 750™ anti-mouse CD4
Brilliant Violet 650™ anti-mouse CD4
Spark Blue™ 550 anti-mouse CD4
Spark NIR™ 685 anti-mouse CD4
KIRAVIA Blue 520™ anti-mouse CD4
PE/Fire™ 640 anti-mouse CD4
APC/Fire™ 810 anti-mouse CD4
PE/Fire™ 700 anti-mouse CD4
Spark Violet™ 538 anti-mouse CD4
Spark YG™ 593 anti-mouse CD4
Spark Blue™ 574 anti-mouse CD4 Antibody
Spark UV™ 387 anti-mouse CD4