14-2444-82

antibody from Invitrogen Antibodies

Targeting: CD4

Western blot (Recommended by provider)

Immunohistochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

14-2444-82

Provider

Invitrogen Antibodies

Product name

CD4 Monoclonal Antibody (N1UG0), eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Description: The N1UG0 monoclonal antibody reacts with human CD4, a 59 kDa cell surface glycoprotein expressed by the majority of thymocytes, a subpopulation of mature T cells (T-helper cells) and in low levels on monocytes. CD4 is a receptor for the human immunodeficiency virus (HIV). The N1UG0 antibody is recommended for use in staining human formalin-fixed paraffin embedded tissue sections. Applications Reported: This N1UG0 antibody has been reported for use in western blotting and immunohistochemical staining of formalin-fixed paraffin embedded tissue sections. Applications Tested: This N1UG0 antibody has been tested by immunohistochemistry of formalin-fixed paraffin embedded human tissue using low or high pH antigen retrieval at less than or equal to 2.5 µg/mL. This N1UG0 antibody has been tested by western blot on reduced PBMC and can be used at less than or equal to 5.0 µg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC. Filtration: 0.2 µm post-manufacturing filtered.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

N1UG0

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

4° C

References

Submitted references

Molecular Mechanism of Sphingosine-1-Phosphate Receptor 1 Regulating CD4(+) Tissue Memory in situ T Cells in Primary Sjogren's Syndrome.

Yang XX, Yang C, Wang L, Zhou YB, Yuan X, Xiang N, Wang YP, Li XM
International journal of general medicine 2021;14:6177-6188

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Highly multiplexed immunofluorescence imaging of human tissues and tumors using t-CyCIF and conventional optical microscopes.

Lin JR, Izar B, Wang S, Yapp C, Mei S, Shah PM, Santagata S, Sorger PK
eLife 2018 Jul 11;7

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Immunofluorescence-detected infiltration of CD4+FOXP3+ regulatory T cells is relevant to the prognosis of patients with endometrial cancer.

Yamagami W, Susumu N, Tanaka H, Hirasawa A, Banno K, Suzuki N, Tsuda H, Tsukazaki K, Aoki D
International journal of gynecological cancer : official journal of the International Gynecological Cancer Society 2011 Dec;21(9):1628-34

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Inhibition of gp160 and CD4 maturation in U937 cells after both defective and productive infections by human immunodeficiency virus type 1.

Bour S, Boulerice F, Wainberg MA
Journal of virology 1991 Dec;65(12):6387-96

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Separation of functional subsets of human T cells by a monoclonal antibody.

Reinherz EL, Kung PC, Goldstein G, Schlossman SF
Proceedings of the National Academy of Sciences of the United States of America 1979 Aug;76(8):4061-5

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry of formalin-fixed paraffin embedded human tonsil using 2.5 µg/mL of Anti-Human CD4 Purified, followed by Anti-Mouse IgG Biotin, Streptavidin HRP, and DAB visualization. Nuclei are counterstained with hematoxylin.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of CD4 was performed using formalin-fixed paraffin-embedded human tonsil tissue sections. To expose the target protein, heat-induced epitope retrieval was performed on de-paraffinized sections using eBioscience™ IHC Antigen Retrieval Solution - High pH (10X) (Product # 00-4956-58) diluted to 1X solution in water in a decloaking chamber at 110 degree Celsius for 15 minutes. Following antigen retrieval, the sections were blocked with 2% normal goat serum in 1X PBS for 45 minutes at room temperature and then probed with or without CD4 Monoclonal Antibody (N1UG0) (Product #14-2444-82) at 5 µg/mL in 0.1% normal goat serum overnight at 4 degree Celsius in a humidified chamber. Detection was performed using Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32723) at a dilution of 1:2000 in 0.1% normal goat serum for 45 minutes at room temperature. ReadyProbes™ Tissue Autofluorescence Quenching Kit (Product # R37630) was used to quench autofluorescence from the tissues. Nuclei were stained with DAPI (Product # D1306) and the sections were mounted using ProLong™ Glass Antifade Mountant (Product # P36984). The images were captured on EVOS™ M7000 Imaging System (Product # AMF7000) at 20X magnification and externally deconvoluted.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of CD4 was performed using formalin-fixed paraffin-embedded human tonsil tissue sections. To expose the target protein, heat-induced epitope retrieval was performed on de-paraffinized sections using eBioscience™ IHC Antigen Retrieval Solution - Low pH (10X) (Product # 00-4955-58) diluted to 1X solution in water in a decloaking chamber at 110 degree Celsius for 15 minutes. Following antigen retrieval, the sections were blocked with 2% normal goat serum in 1X PBS for 45 minutes at room temperature and then probed with or without CD4 Monoclonal Antibody (N1UG0) (Product #14-2444-82) at 5 µg/mL in 0.1% normal goat serum overnight at 4 degree Celsius in a humidified chamber. Detection was performed using Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32723) at a dilution of 1:2000 in 0.1% normal goat serum for 45 minutes at room temperature. ReadyProbes™ Tissue Autofluorescence Quenching Kit (Product # R37630) was used to quench autofluorescence from the tissues. Nuclei were stained with DAPI (Product # D1306) and the sections were mounted using ProLong™ Glass Antifade Mountant (Product # P36984). The images were captured on EVOS™ M7000 Imaging System (Product # AMF7000) at 20X magnification and externally deconvoluted.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

NULL

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

NULL

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 1 S1PR1 expression in CD4 + CD69 + T RM cells. ( A ) Lymphocyte infiltration was detected by H&E staining of the labial salivary glands(LSG) of patients with pSS. ( B ) Expression of S1PR1 was detected by immunohistochemistry in labial salivary glands (LSG) of patients with pSS. ( C ) Immunofluorescence was performed to detect the expression of CD4, CD69 and S1PR1 in LSG samples with lymphocyte infiltration foci. Bar=20mum. DAPI was used to stain the nuclei and glowed blue. CD4 was pink light, CD69 was red light, and S1PR1 was green light. The yellow arrows point to positive fluorescence results and the white arrows point to negative fluorescence results in the DAPI, CD4, CD69 and S1PR1 image. In the in Merge image, the yellow arrows represent only S1PR1 expression in CD4 + T cells and no surface expression of CD69, the white arrows point to the CD4 + CD69 + T cells without the expression of S1PR1.