12-6644-42

antibody from Invitrogen Antibodies

Targeting: TNFRSF10A Apo2, CD261, DR4, TRAILR-1, TRAILR1

Provider product page for 12-6644-42

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [25]
Comments [0]
Validations
Flow cytometry [1]
Other assay [5]

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Product number: 12-6644-42 - Provider product page
Provider: Invitrogen Antibodies
Product name: CD261 (DR4) Monoclonal Antibody (DJR1), PE, eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: The DJR1 monoclonal antibody reacts with human DR4, also known as TRAIL-R1. DR4 binds to TRAIL, activates NF-kappaB, and induces TRAIL-mediated apoptosis. DR4 is expressed by a variety of human tumor cells and at very low level on human leukocytes. Expression has also been found inside the cell on some cell lines (Zhang et al). This has been confirmed with intracellular staining of Jurkat cell line. Please contact tech@ebioscience.com for more information. Applications Reported: The DJR1 antibody has been reported for use in flow cytometric analysis. Applications Tested: This DJR1 antibody has been pre-titrated and tested by flow cytometric analysis of human DR4-transfected cells. This can be used at 5 µL (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. 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: DJR1
Vial size: 100 Tests
Concentration: 5 µL/Test
Storage: 4° C, store in dark, DO NOT FREEZE!

TNFRSF10A protein structure - 12-6644-42 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: 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: 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

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 4 Orthogonal genetic screens and surface marker analysis specify SLC39A7 loss-of-function phenotype on receptor trafficking. a Spider plot summarizing the results of 8 independent CRISPR/ Cas9 screens in KBM7 wildtype, KBM7 FADD - , or HAP1 cells using an SLC-specific gRNA library. Each plot section represents one screen with the indicated stimuli. Screen analysis was performed by identifying differentially enriched sgRNAs using DESeq2 and then aggregating sgRNAs to genes using Gene Set Enrichment Analysis. Identified hits are ranked according to the adjusted p -value of enrichment (-log10(p adj )), bubble size indicates the number of significantly enriched sgRNAs and color the average log2 fold-change (aLFC) of the enriched sgRNAs. Screens were performed in duplicate. SLC39A7 is highlighted in orange. No gene was identified in KBM7 wildtype cells upon TRAIL treatment. b MCA of Jurkat E6.1 SpCas9 cells transduced with a GFP marker (GFP + ) and sgRNAs targeting either SLC39A7 or Renilla luciferase (sg Ren ) as control, against cells transduced with sg Ren and an mCherry marker (mCherry + ). The cell populations were mixed at 1:1 ratio, treated with TRAIL (20 ng/ml) or FASL (1 ng/ml), and analyzed after 14 days by flow cytometry. Data represent mean value +- s.d. of two independent experiments performed in duplicates, n.d. (not determined) indicates wells with no outgrowth. c Flow cytometry analysis for TRAILR1 (left) and TRAILR2 (right) surface expression in KBM7 FADD - ,
Conjugate: Yellow dye