Antibody data
- Antibody Data
- Antigen structure
- References [14]
- Comments [0]
- Validations
- Flow cytometry [1]
- Other assay [14]
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- Product number
- 61-1278-42 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CD127 Monoclonal Antibody (eBioRDR5), PE-eFluor™ 610, eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: The eBioRDR5 monoclonal antibody reacts with human CD127 (Interleukin-7 Receptor alpha). CD127 complexes with CD132, also known as the common gamma chain (gamma c), to form the multi-functional IL-7 receptor (IL-7R). CD127 is a type I glycoprotein with a molecular weight of 75-80 kDa and is expressed by immature B cells through the early pre-B stage, by thymocytes during several stages of their development, and on most mature T cells, with transient down-regulation upon activation. Binding of IL-7 results in signal transduction which occurs through several tyrosine kinase pathways including the Jak/STAT pathway. IL-7 is indispensible for the development of lymphocytes, and the control of homeostatic proliferation of T-cells in the periphery. In addition, IL-7R signaling is know to be involved in the regulation of T cell receptor (TCR) locus rearrangement in gamma delta T cells. Interestingly, recently it has been demonstrated that CD127 expression is down-regulated on CD4+CD25+ regulatory T cells (T regs). While the co-expression of CD4 and CD25 has become widely used as an indicator of T regs, this method of identification may also include cells without suppressive activity. It has clearly been shown that CD4+CD25+ cells that have down-regulated the expression of CD127 are significantly more highly-enriched for the regulatory T population, as defined by expression of the T reg-specific transcription factor Foxp3 and the suppressive activity of these cells, in vitro. Binding of the eBioRDR5 monoclonal antibody to PBMCs is blocked by pre-incubation of the cells with recombinant human IL-7 (Product # 14-1079-80). Applications Reported: This eBioRDR5 antibody has been reported for use in flow cytometric analysis. Applications Tested: This eBioRDR5 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.125 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. PE-eFluor® 610 can be excited with laser lines from 488-561 nm and emits at 607 nm. We recommend using a 610/20 band pass filter (equivalent to PE-Texas Red®). Please make sure that your instrument is capable of detecting this fluorochome. Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 488-561 nm; Emission: 607 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser. Filtration: 0.2 µm post-manufacturing filtered.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- eBioRDR5
- Vial size
- 100 Tests
- Concentration
- 5 µL/Test
- Storage
- 4° C, store in dark, DO NOT FREEZE!
Submitted references Targeting the mTOR pathway uncouples the efficacy and toxicity of PD-1 blockade in renal transplantation.
The Effects of High Mobility Group Box-1 Protein on Peripheral Treg/Th17 Balance in Patients with Atherosclerosis.
Activation of miR-21-Regulated Pathways in Immune Aging Selects against Signatures Characteristic of Memory T Cells.
A probiotic modulates the microbiome and immunity in multiple sclerosis.
Characterization of CD127(-) CD25(++) Treg from human colostrum.
Serial immunomonitoring of cancer patients receiving combined antagonistic anti-CD40 and chemotherapy reveals consistent and cyclical modulation of T cell and dendritic cell parameters.
HDAC inhibition potentiates immunotherapy in triple negative breast cancer.
Blocking the recruitment of naive CD4(+) T cells reverses immunosuppression in breast cancer.
The Distribution of Human Stem Cell-like Memory T Cell in Lung Cancer.
Enhanced suppressor function of TIM-3+ FoxP3+ regulatory T cells.
Fc receptor-like 3 protein expressed on IL-2 nonresponsive subset of human regulatory T cells.
Allosuppressive donor CD4+CD25+ regulatory T cells detach from the graft and circulate in recipients after liver transplantation.
Loss of IL-7 receptor alpha on CD4+ T cells defines terminally differentiated B cell-helping effector T cells in a B cell-rich lymphoid tissue.
Loss of IL-7 receptor alpha on CD4+ T cells defines terminally differentiated B cell-helping effector T cells in a B cell-rich lymphoid tissue.
Esfahani K, Al-Aubodah TA, Thebault P, Lapointe R, Hudson M, Johnson NA, Baran D, Bhulaiga N, Takano T, Cailhier JF, Piccirillo CA, Miller WH
Nature communications 2019 Oct 17;10(1):4712
Nature communications 2019 Oct 17;10(1):4712
The Effects of High Mobility Group Box-1 Protein on Peripheral Treg/Th17 Balance in Patients with Atherosclerosis.
Ding JW, Zhou T, Zheng XX, Wang XA, Tong XH, Luo CY, Zhang ZQ, Yu B
Acta Cardiologica Sinica 2018 Sep;34(5):399-408
Acta Cardiologica Sinica 2018 Sep;34(5):399-408
Activation of miR-21-Regulated Pathways in Immune Aging Selects against Signatures Characteristic of Memory T Cells.
Kim C, Hu B, Jadhav RR, Jin J, Zhang H, Cavanagh MM, Akondy RS, Ahmed R, Weyand CM, Goronzy JJ
Cell reports 2018 Nov 20;25(8):2148-2162.e5
Cell reports 2018 Nov 20;25(8):2148-2162.e5
A probiotic modulates the microbiome and immunity in multiple sclerosis.
Tankou SK, Regev K, Healy BC, Tjon E, Laghi L, Cox LM, Kivisäkk P, Pierre IV, Hrishikesh L, Gandhi R, Cook S, Glanz B, Stankiewicz J, Weiner HL
Annals of neurology 2018 Jun;83(6):1147-1161
Annals of neurology 2018 Jun;83(6):1147-1161
Characterization of CD127(-) CD25(++) Treg from human colostrum.
Cérbulo-Vázquez A, Hernández-Peláez G, Arriaga-Pizano LA, Bautista-Pérez P, Romero-Venado J, Flores-González JC, Figueroa-Damian R, Soriano-Becerril D, Mancilla-Herrera I
American journal of reproductive immunology (New York, N.Y. : 1989) 2018 Feb;79(2)
American journal of reproductive immunology (New York, N.Y. : 1989) 2018 Feb;79(2)
Serial immunomonitoring of cancer patients receiving combined antagonistic anti-CD40 and chemotherapy reveals consistent and cyclical modulation of T cell and dendritic cell parameters.
McDonnell AM, Cook A, Robinson BWS, Lake RA, Nowak AK
BMC cancer 2017 Jun 15;17(1):417
BMC cancer 2017 Jun 15;17(1):417
HDAC inhibition potentiates immunotherapy in triple negative breast cancer.
Terranova-Barberio M, Thomas S, Ali N, Pawlowska N, Park J, Krings G, Rosenblum MD, Budillon A, Munster PN
Oncotarget 2017 Dec 26;8(69):114156-114172
Oncotarget 2017 Dec 26;8(69):114156-114172
Blocking the recruitment of naive CD4(+) T cells reverses immunosuppression in breast cancer.
Su S, Liao J, Liu J, Huang D, He C, Chen F, Yang L, Wu W, Chen J, Lin L, Zeng Y, Ouyang N, Cui X, Yao H, Su F, Huang JD, Lieberman J, Liu Q, Song E
Cell research 2017 Apr;27(4):461-482
Cell research 2017 Apr;27(4):461-482
The Distribution of Human Stem Cell-like Memory T Cell in Lung Cancer.
Hong H, Gu Y, Sheng SY, Lu CG, Zou JY, Wu CY
Journal of immunotherapy (Hagerstown, Md. : 1997) 2016 Jul-Aug;39(6):233-40
Journal of immunotherapy (Hagerstown, Md. : 1997) 2016 Jul-Aug;39(6):233-40
Enhanced suppressor function of TIM-3+ FoxP3+ regulatory T cells.
Gautron AS, Dominguez-Villar M, de Marcken M, Hafler DA
European journal of immunology 2014 Sep;44(9):2703-2711
European journal of immunology 2014 Sep;44(9):2703-2711
Fc receptor-like 3 protein expressed on IL-2 nonresponsive subset of human regulatory T cells.
Nagata S, Ise T, Pastan I
Journal of immunology (Baltimore, Md. : 1950) 2009 Jun 15;182(12):7518-26
Journal of immunology (Baltimore, Md. : 1950) 2009 Jun 15;182(12):7518-26
Allosuppressive donor CD4+CD25+ regulatory T cells detach from the graft and circulate in recipients after liver transplantation.
Demirkiran A, Bosma BM, Kok A, Baan CC, Metselaar HJ, Ijzermans JN, Tilanus HW, Kwekkeboom J, van der Laan LJ
Journal of immunology (Baltimore, Md. : 1950) 2007 May 15;178(10):6066-72
Journal of immunology (Baltimore, Md. : 1950) 2007 May 15;178(10):6066-72
Loss of IL-7 receptor alpha on CD4+ T cells defines terminally differentiated B cell-helping effector T cells in a B cell-rich lymphoid tissue.
Lim HW, Kim CH
Journal of immunology (Baltimore, Md. : 1950) 2007 Dec 1;179(11):7448-56
Journal of immunology (Baltimore, Md. : 1950) 2007 Dec 1;179(11):7448-56
Loss of IL-7 receptor alpha on CD4+ T cells defines terminally differentiated B cell-helping effector T cells in a B cell-rich lymphoid tissue.
Lim HW, Kim CH
Journal of immunology (Baltimore, Md. : 1950) 2007 Dec 1;179(11):7448-56
Journal of immunology (Baltimore, Md. : 1950) 2007 Dec 1;179(11):7448-56
No comments: Submit comment
Supportive validation
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- Invitrogen Antibodies (provider)
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- Experimental details
- Staining of normal human peripheral blood cells with Anti-Human CD3 APC (Product # 17-0036-42) and Mouse IgG1 K Isotype Control PE-eFluor® 610 (Product # 61-4714-82) (left) or Anti-Human CD127 PE-eFluor® 610 (right). Cells in the lymphocyte gate were used for analysis.
Supportive validation
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- FIGURE 1 Identification of Tscm CD4 + CD45RA + CD45RO - CD62L + CCR7 + CD127 + CD27 + CD28 + CD95 + CD122 + T (Tscm) cell in human blood and lymph nodes. PBMCs were isolated from the blood of non-small cell lung cancer (NSCLC) patients (n=15) (NSCLC-PBMC) and healthy donors (n=11) (HD-PBMC); lymphocytes were isolated from the tumor-infiltrated lymph node of NSCLC patients who were collected blood at same time (n=7) (NSCLC-Ly); lymphocytes were isolated from the healthy lymph node of non lung cancer patients (n=7) (Normal-Ly), analyzed by flow cytometry. A, Representative flow cytometric analyses of CD4 + CD45RA + CD45RO - CD62L + CCR7 + CD127 + CD27 + CD28 + CD95 + CD122 + T cells, indicating Tscm cells. B, The frequency of the CD4 + Tscm cells in the HD-PBMC, NSCLC-PBMC, Normal-Ly, NSCLC-Ly. The events of CD4 + Tscm cells in the blood and lymph node from NSCLC patients and healthy donors, expressed as the mean+-SEM. C, The frequency of the CD8 + Tscm cells in the HD-PBMC, NSCLC-PBMC, Normal-Ly, NSCLC-Ly, expressed as the mean+-SEM. D, The events of Tscm of CD4 + and CD8 + cells in the blood and the lymph node from NSCLC patients and healthy donors. HD indicates healthy donors; IFN, interferon; PBMC, peripheral blood mononuclear cells; Tscm cell, stem cell-like memory T cell. * P
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- FIGURE 3 CD4 + CD45RA + CD45RO - CD95 - CD122 + CD127 + T cell population displays different phenotypes in human blood and lymph nodes. A, Flow cytometry plots showing IFN-gamma expression in the CD4 + CD45RA + /CD45RO - CD95 - CD122 + CD127 + T cells from the blood and lymph node of the non-small cell lung cancer (NSCLC) patients and healthy donors. B, The mean frequency (+-SEM) of the events of CD4 + CD45RA + /CD45RO - CD95 - CD122 + CD127 + T cells. C, The mean IFN-gamma production (+-SEM) of the CD4 + CD45RA + /CD45RO - CD95 - CD122 + CD127 + T cells. HD indicates healthy donors; IFN, interferon; PBMC, peripheral blood mononuclear cells; PMA, phorbol 12-myristate13-acetate. * P
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- Fig. 6 a Representative flow cytometry data, demonstrating the gating strategy used on PBMC for Treg identification and analysis. FSC-area vs. FSC-height was used for doublet discrimination. A ""dump"" channel was used to gate out dead cells (LIVE/DEAD fixable viability stain), CD14 + monocytes and CD19 + B cells, and lymphocytes were subsequently selected by FSC vs. SSC. CD4 + T cells were gated on the basis of CD4 vs. CD3 staining, then examined for expression of Ki67 and ICOS. Tregs were identified within the CD4 + T cell population as CD25 hi CD127 lo and Foxp3 + . b Longitudinal empirical data, linear mixed models and estimated means (left, centre and right-hand panels respectively) for Ki67+ and ICOS+ expression on CD4+ T cells, and the Treg proportion of CD4 cells ( P -values: *
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- Figure 6 In vivo knockdown of PITPNM3 in CD4 + T cells reverses immunosuppression and inhibits tumor progression in humanized mice. (A) Humanized mice bearing palpable MDA-MB-231 orthotopic xenografts were intraperitoneally injected daily for 14 days with PBS, 1 nmol CD4-aptamer-control siRNA (AsiC-con) or CD4-aptamer-siRNA targeting PITPNM3 (sequence in A , AsiC-PI) to assess the role of PITPNM3 in TI Tregs, and other T cells and tumor control. Experimental schematic is provided in Supplementary information, Figure S9A . (B) Representative immunoblots showing selective knockdown of PITPNM3 protein in PB CD4 + T cells, but not tumor xenografts ( n = 3). (C) PITPNM3 knockdown did not affect the distribution of human CD45 + hematopoietic cells, CD4 + and CD8 + T cells, and CD14 + monocytes in the peripheral blood of humanized mice. Representative flow plots are shown ( n = 3). (D , E) Effect of PITPNM3 knockdown on TI naive CD4 + , Tregs and CD8 + T cell numbers, and apoptosis by TUNEL assay in xenografts. D shows representative immunofluorescence microscopy images. Top row indicates CD4 + naive T cells by arrows; the second row indicates CD4 + CD45RO + Foxp3 - CD4 + memory T cells (yellow arrows) and Foxp3 + Tregs (white arrows). Scale bar, 50 mum. E shows number of cells of each subtype/high power field in eight mice ( ** P < 0.01, *** P < 0.001 compared to PBS group by Student's t -test). (F) Flow cytometry analysis of gated human CD3 + CD4 + cells isolated from xenogra
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- Figure 4. miR-21 Promotes Effector Cell Differentiation through AP-1 Activation (A) Naive CD4 + T cells were transfected with either scrambled control or miR-21-blocking locked nucleic acid (LNA21). After 48 hr, PDCD4 and b-actin expression were assessed by western blot. Representative blots and mean normalized intensities from four experiments are shown (mean +- SEM, two-tailed paired t test). (B) Naive CD4 + T cells were activated with anti-CD3 and anti-CD28 beads and transduced with a lentiviral vector expressing scrambled control or anti-miR-21. The representative histogram shows phosphorylated JNK in GFP + cells on day 3. The filled gray histogram represents unstimulated naive CD4 + T cells. Results from 7 experiments are expressed relative to the geometric mean fluorescence intensity (MFI) of controls (mean +- SEM, two-tailed paired t test). (C) Naive CD4 + T cells were activated and transduced as described in (B) and co-transfected with the AP-1 luciferase reporter plasmid and the Renilla luciferase control construct. On day 3, the activity of AP-1 firefly luciferase was measured and normalized to that of Renilla luciferase (n = 4, mean +- SEM, two-tailed paired t test). (D-F) Naive CD4 + T cells isolated from 20- to 35-year-old and 65- to 85-year-old individuals were activated with beads coated with anti-CD3 and anti-CD28 antibodies. (D) PDCD4 expression was quantified by RT-PCR on day 3 and day 5. Results are normalized to ACTB and presented relative to