12-6988-80

antibody from Invitrogen Antibodies

Targeting: RORC NR1F3, RORG, RZRG, TOR

Provider product page for 12-6988-80

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [106]
Comments [0]
Validations
Flow cytometry [1]
Other assay [15]

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Product number: 12-6988-80 - Provider product page
Provider: Invitrogen Antibodies
Product name: ROR gamma (t) Monoclonal Antibody (AFKJS-9), PE, eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: The AFKJS-9 monoclonal antibody reacts with the mouse, human and rhesus monkey ROR gamma (t) protein. ROR gamma is a member of the retinoic acid-related orphan receptor (ROR) family, which also includes ROR alpha and ROR beta. ROR family proteins are ligand-dependent transcription factors that play roles in multiple physiological processes. ROR gamma is expressed in several tissues including liver, lung, muscle, heart and kidney. Furthermore, it was discovered that alternative transcription results in the expression of an isoform, ROR gamma (t), which is expressed exclusively in cells of the lymphoid compartment, namely CD4+CD8+ "double-positive" thymocytes, Th17 cells of the periphery and lymphoid tissue inducer (LTi) cells of lymphoid organs. The ROR gamma (t) isoform differs from ROR gamma by three unique amino acids at its amino terminus. Therefore, the AFKJS-9 antibody will react with both the ROR gamma and ROR gamma (t) isoforms. Applications Reported: This AFKJS-9 antibody has been reported for use in intracellular staining followed by flow cytometric analysis. Applications Tested: This AFKJS-9 antibody has been tested by intracellular staining followed by flow cytometric analysis of mouse thymocytes and Th17-polarized splenocytes using the Foxp3/Transcription Factor Staining Buffer Set (Product # 00-5523-00) and protocol. Please refer to Best Protocols: Protocol B: One step protocol for (nuclear) intracellular proteins located under the Resources Tab online. This can be used at less than or equal to 0.5 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. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Excitation: 488-561 nm; Emission: 578 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser. Filtration: 0.2 µm post-manufacturing filtered.
Reactivity: Human, Mouse
Host: Rat
Conjugate: Yellow dye
Isotype: IgG
Antibody clone number: AFKJS-9
Vial size: 25 µg
Concentration: 0.2 mg/mL
Storage: 4° C, store in dark, DO NOT FREEZE!

RORC protein structure - 12-6988-80 shown in red.

Supportive validation

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Experimental details: NULL
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Yellow dye

Supportive validation

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Experimental details: NULL
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 6 Splenic Th17 cells and pro-inflammatory cytokine expression. The lymphocytes were isolated from mice subjected to various treatments. (A) Flow cytometry was used to evaluate RORgammat + CD3 + CD4 + Th17 cells among splenic CD4 + cells obtained from mice. Quantification of (B) RORgamma and (C) IL-17 from the flow cytometry results. (D) The mRNA levels of the cytokines IL-6, IFN-gamma, TNF-alpha, IL-1beta, and MCP-1beta in the spleen tissue samples were assessed. (E) IL-17 mRNA and protein levels were evaluated by reverse transcription-quantitative polymerase chain reaction, western blot and ELISA. Values are expressed as the mean +- standard error of the mean (n=10 in each group). *** P
Conjugate: Yellow dye

Supportive validation

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Experimental details: Figure 4. Flow cytometric mapping of the expression of forkhead box P3 and retinoic acid-related orphan receptor gammat (ROR-gammat) molecules in CD4 + cells of peripheral blood.
Conjugate: Yellow dye

Supportive validation

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Experimental details: Figure 3 Interleukin (IL)-22 expression is reduced in Citrobacter rodentium -infected CX 3 CR1 GFP/GFP animals. ( a ) From the proximal colon of noninfected and C. rodentium -infected CX 3 CR1 GFP/GFP and wild-type littermate controls mRNA was isolated. cDNA was prepared by reverse transcription and quantitative real-time PCR was performed with cyber green and the indicated primers. beta-actin was used as a housekeeping gene to normalize cDNA input between samples. Normalized ct-values of the untreated samples (baseline) are set to 1 and values are plotted as fold expression of the baseline. The assays were carried out in triplicates. As per indicated, group 4 animals were analyzed. ( b ) IL-22 expression by colonic lamina propria (cLP) cell isolates from noninfected and C. rodentium -infected B6, (age- and sex-matched) and CX 3 CR1 GFP/+ and CX 3 CR1 GFP/GFP animals was analyzed by multicolor flow cytometry. Numbers indicate the percentage of IL-22-positive cells. ( c ) cLP IL22 + cells from noninfected and C. rodentium- infected B6, (age- and sex-matched), and CX 3 CR1 GFP/+ and CX 3 CR1 GFP/GFP animals were stained for CD3e and CD4. Numbers indicate the percentage of CD3e + , CD3e + CD4 + , CD4 + , and CD3e - CD4 - cells within the IL-22 + cell population. Data from an individual representative mouse per group (of five to seven individual mice analyzed per group) are shown. ( d ) cLP IL-22 + CD3e - CD4 + cells were (intracellular) stained for related orphan receptor gamma-
Conjugate: Yellow dye

Supportive validation

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Experimental details: Figure 2 Th17 cell increased in the peripheral blood and the spleen following pCR3.1-cyp26a1 immunization during mice peri-implantation, especially on D5 of mice pregnancy. (A) Flow cytometry scatterplot of Th17 cells. Th17 cells are CD4 + RORgammat + . (B) The Th17 cell proportion increased significantly in the peripheral blood and the spleen under the influence of recombinant plasmid, especially on D5 of pregnancy. The CD4 + RORgammat + Th17 cell ratio was evaluated by flow cytometry analysis. The data are expressed as the% of CD4 + RORgammat + double positive cells. The bars represent the SD of the mean. A paired t -test was used to assess the significance of differences. Pairwise comparisons between each treatment group, * P < 0.05. At least, three independent experiments were repeated for each time-point. A total of 49 samples from pregnant mice were assessed.
Conjugate: Yellow dye

Supportive validation

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Experimental details: Figure 4 Further certainty, the regulation of cyp26a1 on Th17 cells at uterine implantation sites by uterine horn injection of anti-cyp26a1 antibody during mice peri-implantation. (A) Decrease in cyp26a1 levels at D5 uterine implantation sites following uterine horn injection of cyp26a1 antibody on D3 of pregnancy. Cyp26a1 levels were reduced at uterine implantation sites based on Western blotting and immunohistochemistry. beta-actin was used as a loading control. The bars represent the SD of the mean of the relative value in grey (cyp26a1/beta-actin). A paired t -test was used to assess the significance of differences. Pairwise comparisons between each treatment group, * P < 0.05, ** P < 0.01; bars = 50 mum. The data were derived from three separate samples from pregnant mice. At least, three independent experiments were repeated for this time-point. A total of nine samples from pregnant mice were assessed. (B) Th17cell increased in the peripheral blood and the spleen following uterine horn injection of cyp26a1 antibody. Flow cytometry histogram of Th17 cells is on the left side. Th17 cells are CD4 + RORgammat + . Th17 cell ratio was evaluated by flow cytometry analysis. The data are expressed as the% of CD4 + RORgammat + double positive cells. The bars represent the SD of the mean. A paired t -test was used to assess the significance of differences. Pairwise comparisons between each treatment group, * P < 0.05. Three independent experiments were repeated for this time-point
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 Further certainty, the regulation of cyp26a1 on Th17 cells at uterine implantation sites by tail vein injection of cyp26a1 lentiviral interference segment (No. 306) during mice peri-implantation. (A) Decrease in cyp26a1 levels at uterine implantation sites of D5 following tail vein injection of cyp26a1 lentiviral interference segment (306) on D3 of pregnancy. Cyp26a1 levels were reduced at uterine implantation sites based on Western blotting and immunohistochemistry; bars = 25 mum. The data were derived from two separate samples from pregnant mice. At least, three independent experiments were repeated for this time-point. A total of six samples from pregnant mice were assessed. (B) Th17 cell increased in the peripheral blood, but the spleen following tail vein injection of cyp26a1 lentiviral interference segment (No. 306). Flow cytometry histogram of Th17 cells is on the left side. Th17 cells are CD4 + RORgammat + . Th17 cell ratio was evaluated by flow cytometry analysis. The data are expressed as the% of CD4 + RORgammat + double positive cells. The bars represent the SD of the mean. A paired t -test was used to assess the significance of differences. Pairwise comparisons between each treatment group, * P < 0.05. Three independent experiments were repeated. A total of nine samples from pregnant mice were assessed. (C) Th17 levels were significantly reduced at uterine implantation sites based on Western blotting of D5 pregnancy. beta-actin was used as a loading contr
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2. Th17-polarized CD4 + T cells are sufficient to promote hippocampal precursor cell proliferation in adult TCRalpha -/- mice. ( A - C ) Th17 polarization in vitro . ( A ) Flow cytometry of encephalitogenic CD4 + T cells. Dot plots show pre-sort (left) and post-sort (right) analysis of naive T cells (CD4 + CD62L high CD25 - Foxp3 GFP- ) from pooled spleen and lymph nodes of 2D2 x Foxp3 GFP mice. FACS-purified T cell populations were cultured under Th17-polarizing conditions, as described in the Methods section. On day 7, efficiency of Th17 cell differentiation was confirmed by intracellular flow cytometry of ( B ) the Th17 transcription factor ROR-gammat and ( C ) the Th17 and Th1 signature cytokines IL-17 and IFN-gamma, respectively. ( D - I ) Impact of adoptive Th17 cell transfer on hippocampal precursor cell proliferation in TCRalpha -/- mice. ( D ) On day 7, 4 x 10 6 total cells from Th17 polarization cultures were injected i.v. into adult TCRalpha -/- mice. (D1) Dot plots show representative flow cytometry of CD4 + T cells in peripheral blood of recipient mice that express the Valpha3.2 subunit of the transgenic 2D2 TCR, two weeks after adoptive transfer. (D2) and (D3) Graphs show composite percentages of total CD4 + T cells (D2) and MOG 35-55 -reactive Valpha3.2 + T cells among gated CD4 + T cell populations (D3) from peripheral blood of recipient mice. The arrowheads in (D2) and (D3) highlight an individual mouse that exhibited immune cell infiltrations in the b
Conjugate: Yellow dye