12-9908-41

antibody from Invitrogen Antibodies

Targeting: TNFRSF10B CD262, DR5, KILLER, TRAIL-R2, TRAILR2, TRICK2A, TRICKB

Provider product page for 12-9908-41

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [41]
Comments [0]
Validations
Flow cytometry [1]
Other assay [8]

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Product number: 12-9908-41 - Provider product page
Provider: Invitrogen Antibodies
Product name: CD262 (DR5) Monoclonal Antibody (DJR2-4 (7-8)), PE, eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: The DJR2-4 monoclonal antibody reacts with human DR5, also known as TRAIL-R2, Apo2, TRICK2 and KILLER. DR5 binds to TRAIL, activates NF-kappaB, and induces TRAIL-mediated apoptosis. DR5 is expressed broadly by normal tissues as well as several tumor cells. Applications Reported: The DJR2-4 (a.k.a. 7-8) antibody has been reported for use in flow cytometric analysis. Applications Tested: This DJR2-4 (a.k.a. 7-8) antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.25 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. Excitation: 488-561 nm; Emission: 578 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser. Filtration: 0.2 µm post-manufacturing filtered.
Reactivity: Human
Host: Mouse
Conjugate: Yellow dye
Isotype: IgG
Antibody clone number: DJR2-4 (7-8)
Vial size: 25 Tests
Concentration: 5 µL/Test
Storage: 4° C, store in dark, DO NOT FREEZE!

TNFRSF10B protein structure - 12-9908-41 shown in red.

Supportive validation

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

Supportive validation

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

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 BT20 and MCF7 spheroids have a lower expression of death receptors. (A) Confocal micrographs of BT20 and MCF7 cells cultured as 3D spheroids and 2D monolayer stained with antibodies against DR4 (red), DR5 (red) and EpCAM (green). (Scale bar = 20 um) (B) Flow cytometry histograms comparing DR4 and DR5 expression in BT20 and MCF7 spheroids and monolayer. (C) Western blot analysis of total DR4 and DR5 expression in cell lysates from BT20 and MCF7 spheroids and monolayer.
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 3 YIPF2 enhances TNFRSF10B recycling to plasma membrane. a Overexpression of YIPF2 in A549 and H1792 cells. The surface expression of TNFRSF10B was confirmed by flow cytometry analyses. b Knockdown of YIPF2 expression by YIPF2-1 and YIPF2-2 siRNA in A549 cells. The surface expression of TNFRSF10B was confirmed by flow cytometry analyses. c Relative RT-qPCR analyses of YIPF2 and TNFRSF10B mRNA levels after YIPF2 overexpression in H1792 (left) and H1299 (right) cells ( n = 3). d Relative RT-qPCR analyses of YIPF2 and TNFRSF10B mRNA levels after YIPF2 knocking down in H1792 (left) and H1299 (right) cells ( n = 3). e Left: Overexpression of YIPF2 in H1299 cells in the presence or absence of cycloheximide (CHX) at 10 mug/mL for the indicated times (0, 4, 8 and 12 h). Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B and ACTB. Right: The band intensity of TNFRSF10B was quantified by ImageJ software and plotted. This experiment was repeated three times independently with similar results. f Left: Knockdown of YIPF2 expression by YIPF2-1 siRNA in A549 cells in the presence or absence of cycloheximide (CHX) at 10 mug/mL for the indicated times (0, 4, 8 and 12 h). Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B and ACTB. Right: The band intensity of TNFRSF10B was quantified by ImageJ software and plotted. This experiment was repeated three times independently with similar results. (mean +- SEM, n = 3 inde
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 3 RALB silencing regulates DR5 expression levels. A Top Left: HCT116 cells were transfected with 10 nM SC, siRALA, siRALB or siRALGDS for 24 h, following which cells were harvested for protein extraction. Protein was subsequently analysed using a human apoptosis array. Densitometry was performed on the array panels using ImageJ software, and results for DR4, DR5, Fas/CD95, FADD and TNFR1/TNFRSF1A are shown. Top right: PARP, DR5, DR4, FADD, XIAP, Bcl-xL, RALA and RALB levels in CRC cells following RALA, RALB and RALGDS silencing for the indicated times. Bottom left: Wheat germ agglutinin (WGA), a lectin that binds N -acetylglucosamine post-translational modifications on membrane receptors, was used as a plasma membrane marker. Fixed cells were stained with Alexa488-WGA prior to permeabilisation to minimise the staining of glycosylated proteins in the Golgi apparatus. A single confocal image was collected of the Alexa488-WGA stained membrane (green) alongside immunostained endogenous DR5 (red). Laser settings were kept constant between the samples to enable the comparison of membrane-associated DR5 staining in control (SC) and siRALB-treated cells. Bottom right: >20 cells were scored for intensity in ImageJ. The intensity was normalised to cellular area and plotted. The data are representative of three independent experiments. B CRC cells were transfected with 10 nM SC or siRALB for 24 h and DR5 cell-membrane expression was assessed by flow cytometry using a DR5-specific p
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 Protein expression of FADD, IAPs, and TRAILR2/DR5 in HPV(+) HNSCC cell lines. ( A ). Protein expression from whole cell lysates of HOK and a panel of HPV(+) HNSCC cell lines were examined by Western blot. Protein expression was quantified by comparison to HOK and actin as a loading control. ( B ) UM-SCC-47 and UPCI-SCC-90 cells were stained with fluorescent conjugated anti-TRAILR2/DR5 antibody and Zombie-violet for viability. The x-axis represents TRAILR2/DR5 fluorescent intensity, and the y-axis represents cell count. Percentages of TRAILR2/DR5 positivity among live cells for each cell line are represented and compared with the negative controls of unstained cells. ( C ) TRAILR2/DR5 staining of Jurkat cells was served as a positive control for data presented by ( B ), and the unstained cells served as the negative control. ( D ) There was no statistical difference in the percentages of TRAILR2/DR5 positive staining cells between the two HNSCC cell lines. The data are presented by six replicates from two independent experiments. * or ** indicates Student''s t test, p < 0.05 or 0.01.
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 6 COX-2 inhibitor treatment upregulates the expression of DR4 and DR5 in BT20 and MCF7 spheroids. (A) Flow cytometry histograms showing the effect of 100 uM NS-398 treatment on death receptor expression in BT20 and MCF7 cells cultured as spheroids. (B) Western blot analysis of whole cell lysates from untreated and NS-398 treated BT20 and MCF7 spheroids for DR4 and DR5 expression. (C) Flow cytometry scatter plots showing the effect of NS-398 treatment on the prevalence of CD44 hi CD24 lo population in BT20 and MCF7 cells cultured as spheroids.
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 8 COX-2 knockdown reverses TRAIL-resistance in MCF7 spheroid and monolayer cells. (A) qPCR and western blot data showing the relative expression of COX-2 gene (n = 3) and COX-2 protein in control siRNA and COX-2 siRNA transfected MCF7 cells. (B) Flow cytometry histograms and (C) western blot analysis comparing the expression of DR4 in MCF7 cells transfected with control siRNA and COX-2 siRNA cultured as monolayer and spheroids. (D) Bright field images and (E) MTT assay results (n = 5) quantifying the effect of 200 ng/mL of TRAIL in control siRNA and COX-2 siRNA transfected cells cultured as monolayer and spheroids.
Conjugate: Yellow dye