Anti-CD3 epsilon antibody

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody (AB5690)

IHC image of CD3 staining in a formalin fixed, paraffin embedded normal human tonsil tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6) for 20 mins. The section was then incubated with ab5690 at 1/100 dilution for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody (AB5690)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labelling CD3 with ab5690 at 2μl/ml. Slides were steamed in IHC epitope retrieval solution for 35 minutes and then cooled for 20 minutes. Samples were incubated with the primary antibody at room temperature for 1 hour, incubated with a biotinylated secondary antibody for 30 minutes followed by HRP-Streptavidin for 30 minutes. Developed with DAB chromogen substrate for 5-10 minutes. Counter stained with hematoxylin. Magnification : 10X.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody (AB5690)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human skeletal muscle tissue showing no expression of CD3 when labelled with ab5690 at 2μl/ml. Slides were steamed in IHC epitope retrieval solution for 35 minutes and then cooled for 20 minutes. Samples were incubated with the primary antibody at room temperature for 1 hour, incubated with a biotinylated secondary antibody for 30 minutes followed by HRP-Streptavidin for 30 minutes. Developed with DAB chromogen substrate for 5-10 minutes. Counter stained with hematoxylin. Magnification : 40X.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody (AB5690)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labelling CD3 with ab5690 at 2μl/ml. Slides were steamed in IHC epitope retrieval solution for 35 minutes and then cooled for 20 minutes. Samples were incubated with the primary antibody at room temperature for 1 hour, incubated with a biotinylated secondary antibody for 30 minutes followed by HRP-Streptavidin for 30 minutes. Developed with DAB chromogen substrate for 5-10 minutes. Counter stained with hematoxylin. Magnification : 40X.

• IHC-P

AbReview24369****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody (AB5690)

ab5690 staining CD3 epsilon in human lymph node tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded tissue sections). The sections were fixed in paraformaldehyde and subjected to heat-mediated antigen retrieval in citric buffer pH 6.0, prior to blocking with 10% serum for 1 hour at 20°C. The primary antibody was diluted 1/100 and incubated with the sample for 12 hours at 4°C. An HRP-conjugated goat anti-rabbit polyclonal was used as the secondary antibody, diluted 1/200.

This image is courtesy of an Anonymous Abreview.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody (AB5690)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse spleen tissue labelling CD3 with ab5690 at 2μl/ml. Slides were steamed in IHC epitope retrieval solution for 35 minutes and then cooled for 20 minutes. Samples were incubated with the primary antibody at room temperature for 1 hour, incubated with a biotinylated secondary antibody for 30 minutes followed by HRP-Streptavidin for 30 minutes. Developed with DAB chromogen substrate for 5-10 minutes. Counter stained with hematoxylin. Magnification : 40X.

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody (AB5690)

CD3 and active Caspase 3 populations 72 hrs after mouse aortic allograft

C57Bl/6 donor aortic allografts were transplanted into Balb/C recipient mice (N=3 per treatment) and followed up at 72hrs. Compared to saline, Serp-2 but not CrmA treatment reduced caspase 3 activity (panels A-C; p<0.0224). Neither protein treatment significantly reduced CD3+ T cells (panels D-F).

Viswanathan, K. et al PLoS One. 2012;7(9):e44694. doi: 10.1371/journal.pone.0044694. Epub 2012 Sep 26 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody (AB5690)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse spleen tissue labelling CD3 with ab5690 at 2μl/ml. Slides were steamed in IHC epitope retrieval solution for 35 minutes and then cooled for 20 minutes. Samples were incubated with the primary antibody at room temperature for 1 hour, incubated with a biotinylated secondary antibody for 30 minutes followed by HRP-Streptavidin for 30 minutes. Developed with DAB chromogen substrate for 5-10 minutes. Counter stained with hematoxylin. Magnification : 10X.

• WB

Lab

Western blot - Anti-CD3 epsilon antibody (AB5690)

Lanes 1 - 6 : Merged signal (red and green). Green – ab5690 observed at 23 kDa. Red - loading control, ab8245, observed at 37 kDa.

This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab5690 and ab8245 (loading control) overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) at a 1 : 10000 dilution for 1hr at room temperature and then imaged.

All lanes:

Western blot - Anti-CD3 epsilon antibody (ab5690) at 1 µg/mL

Lane 1:

THP1 whole cell lysate (-ve control) at 15 µg

Lane 2:

Raji whole cell lysate (-ve control) at 15 µg

Lane 3:

Jurkat whole cell lysate at 15 µg

Lane 4:

Human Thymus tissue lysate at 15 µg

Lane 5:

Mouse Thymus tissue lysate at 15 µg

Lane 6:

Rat Thymus tissue lysate at 15 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 19 kDa

Observed band size: 23 kDa

false

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody (AB5690)

T cells in central nervous system during late disseminated infection

A-E) Representative epifluorescence images of T cells (CD3), blood vessels (CD31), and nucleated cells (DAPI) in the brain, dura mater, and pia mater. (A) Epifluorescence images described from left to right. CD3 shown in FITC channel; CD31+ blood vessels shown in TRITC channel; nucleated cells shown in DAPI channel; merged image showing CD3+ cell associated with pia mater within the commissure of the isocortex. (B) T cells within the lymphatic-like vascular region of the sagittal sinus in the dura mater. (C) T cell associated with a blood vessel in the vasculature of the brain choroid plexus. (D) T cell associated with blood vessel in the dura mater. (E) T cell in extravascular region of the dura mater.

(After Figure 3 of Dirvan et al)

Divan, A. et al LoS One. 2018 May 3;13(5):e0196893. doi: 10.1371/journal.pone.0196893. eCollection 2018 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

• WB

Supplier Data

Western blot - Anti-CD3 epsilon antibody (AB5690)

Lanes 1 - 2:

Western blot - Anti-CD3 epsilon antibody (ab5690) at 1 µg/mL

Lanes 3 - 4:

No primary antibody

Lanes 1 and 3:

Jurkat cell lysate at 30 µg

Lanes 2 and 4:

Rat thymus tissue lysate at 20 µg

Secondary

All lanes:

Goat anti-rabbit IgG (H+L), highly cross - adsorbed, HiLyte™ Fluor 750-labeled at 1/12500 dilution

Predicted band size: 19 kDa

Observed band size: 23 kDa

false

• IHC

CiteAb

Immunohistochemistry - Anti-CD3 epsilon antibody (AB5690)

CD3 epsilon immunohistochemistry using Anti-CD3 epsilon antibody, ab5690. Publication image from Ng, K. W. et al., 2023, Nature, PubMed : 37046094.

B cell responses in mouse LUAD.a, Immunostaining of B220 (B cells), CD3 (T cells) and TTF1 (tumour cells) in lungs from mice bearing KPAR tumours (scale bars, 500 µm). Representative images of five mice. b, B220 and CD3 immunofluorescence and DAPI staining in KPAR tumour-bearing lungs (scale bars, 20 µm). Representative images of six mice. c, Quantification of PNA+ mature TLS and GCs by histochemistry in KPB6 (n = 10) and KPAR (n = 4) tumour-bearing lung lobes. d, Flow cytometry quantification of B220+GL7+CD95+ GC B cells and TCRβ+CD4+PD-1+CXCR5+ TFH cells in naive and KPAR tumour-bearing lungs (n = 12 mice per group from three experiments). e, Time-course quantification by flow cytometry of B220+EYFP+ and TFH cells in KPAR lungs and draining lymph nodes (dLNs) from AicdaCreERT2Rosa26LSL-EYFP mice (n = 6 mice per time point from one experiment). f, Time-course quantification of KPAR-binding IgM, IgG and IgA from KPAR serum (n = 6). Dashed lines denote the mean staining intensity of naive serum. MFI, mean fluorescence intensity. g, Survival of KPAR recipient mice treated with pooled serum from KPAR tumour-bearing or naive donor mice (n = 12 mice per group from two experiments). h, Representative images (scale bars, 50 µm) and quantification of intratumoural NCR1+ NK cells in KPAR recipients that were untreated or treated with naive or KPAR serum (n = 8 mice per group from two experiments). i, Flow cytometry quantification of NK1.1+CD16+ NK cells in lungs of KPAR recipients that were untreated or treated with naive or KPAR serum (n = 6 mice per group). j, Survival of KPAR recipient mice treated with naive serum (n = 14) or with KPAR serum and anti-NK1.1 (n = 6), anti-CD8 (n = 8) or isotype control (n = 14) (from two experiments). Data in c–f,h,i are represented as mean ± s.e.m. P values were calculated by two-sided Mann–Whitney rank-sum test in c and d (left), two-sided Student’s t test in d (right), one-way ANOVA with Bonferroni correction for multiple comparisons in h,i and log-rank test in g,j.Source data

• WB

CiteAb

Western blot - Anti-CD3 epsilon antibody (AB5690)

CD3 epsilon western blot using anti-CD3 epsilon antibody ab5690. Publication image and figure legend from Guido, M. C., Marques, A. F., et al., 2017, Oxid Med Cell Longev, PubMed 28781721.

ab5690 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab5690 please see the product overview.

(a) Total inflammatory cell count in SEN and INT areas. HE-stained sections, under 400x magnification. θp < 0.05 versus the DM SEN area; *p < 0.01 versus the CT SEN area, #p < 0.01 versus the DM SEN area; †p < 0.01 versus the CT INT area. Data expressed in mean ± SEM. Western blot protein expression analysis depicting CD68 (b), CD3 (c), MCP-1 (d), TNF-α (e), IL1-β (f), and IL-6 (g) was performed 8 weeks after diabetes induction. *p < 0.05 and †p < 0.001 versus the CT group. Data expressed in mean ± SEM in all plots.

false