63-1338-42

antibody from Invitrogen Antibodies

Targeting: PROM1 AC133, CD133, CORD12, MCDR2, PROML1, RP41, STGD4

Provider product page for 63-1338-42

Flow cytometry

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Antibody data

Antibody Data
Antigen structure
References [6]
Comments [0]
Validations
Flow cytometry [1]
Other assay [10]

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Product number: 63-1338-42 - Provider product page
Provider: Invitrogen Antibodies
Product name: CD133 (Prominin-1) Monoclonal Antibody (TMP4), Super Bright™ 600, eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: The TMP4 monoclonal antibody reacts with human CD133 (Prominin-1), a 120 kDa member of the pentaspan family of proteins, which also includes Prominin-2. Their expression is found within plasma membrane protrusions such as epithelial microvilli. CD133 can exist in a number of alternatively spliced isoforms, and the protein has several N-linked glycosylation sites: the occurrence of both may be tissue-dependent. Human CD133 was first identified as an epitope expressed on CD34+ hematopoietic progenitors. Although the ligand and function of CD133 remain unknown, it has since proven to be very useful as a marker for both stem cells and cancer stem cells. In addition to its expression on hematopoietic precursors, CD133 has been used to identify tumorigenic colon cancer stem cells, brain cancer stem cells, prostate cancer stem cells, in addition to others. The binding of the TMP4 antibody does not block the binding of another anti-human CD133 antibody, EMK08 (Product # 12-1339) indicating that they recognize distinct epitopes. This TMP4 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. Super Bright 600 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 600 nm. We recommend using a 610/20 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome. When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information. Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. 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 2-8°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorochrome performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 405 nm; Emission: 600 nm; Laser: Violet Laser Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: TMP4
Vial size: 100 Tests
Concentration: 5 µL/Test
Storage: 4° C, store in dark, DO NOT FREEZE!

PROM1 protein structure - 63-1338-42 shown in red.

Supportive validation

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

Supportive validation

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

Supportive validation

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

Supportive validation

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

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 2 Immunofluorescence staining for detection of malignant cells in the salvaged blood. Blood was collected and washed with an automated salvaging device. Nucleated cells were stained with CK19-PE (white arrow) and CD133-FITC (yellow arrow) (A). Positive cells were eliminated after M-LDF treatment (B).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig 6 Identification of adherent cells with immunofluorescence staining. Cell suspensions were prepared by adding 8.74x10 7 tumor cells from the same patient to the salvaged blood. Adherent cells were stained with AE1/AE3-PE and CD133-FITC. Adherent cells (both AE1/AE3 + and CD133 + ) that were characterized by invasive growth were found in 85% of samples (A), but none after the M-LDF treatment (B).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 1 Identification of EPCs. a Adherent cells grew in a blood island manner. Fluorescent staining of EPCs. b Adherent cells took up UEA-1-lectin. c Adherent cells took up Dil-Ac-LDL. d Adherent cells took up UEA-1-lectin and Dil-Ac-LDL. E. Surface molecular markers of EPCs. Adherent cells expressed CD34, CD133, CD14 and VEGFR-2. All experiments involving cell culture studies were repeated three times with three replicates per experiment

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 Stemness markers of monolayers and spheroids from RT112 and 5637 cells. ( a - c ) Median fluorescence intensity of Aldefluor TM ( a ), CD44 ( b ) and CD133 ( c ) by flow cytometry analysis on RT112 and 5637 cells grown as monolayers. Results are the mean of two ( a ) and three ( b , c ) experimental replicates. Statistical test: t -test, * for p < 0.05. ( d ) Representative images from confocal immunofluorescence (IF) microscopy of RT112 and 5637 cells using SOX2 Antibody (red) and Hoechst 33342 (blue) for nuclei. Cells were grown as monolayer or spheroids on different (Tissue culture-treated, Not-treated or Cell repellent) supports, before being seeded in adherent condition on chamber slides for IF.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 USP4 promotes lung cancer cell stemness and its high expression is correlated with human lung cancer patients. ( A ) The Oncomine dataset ""Bhattacharjee Lung"" was used to analyze Pearson correlation of USP4 and Oct4/Sox2 expression. ( B - E ) H1975 cells stably expressing shRNA against USP4 (shUSP4-#1 or shUSP4-#2) were subjected to ( B ) Western blot analyses, ( C - D ) FACS analyses for CD133-stained cells or ( E ) tumorsphere formation assay. Respective images and quantitation were shown. Data from three independent experiments in triplicates were presented as means +- SD. *** p < 0.001. Scale bar = 100 mum. ( F - I ) H1975 cells stably expressing Flag-USP4 or Flag-USP4 C311A were subjected to ( F ) Western blot analyses, ( G ) FACS analyses for CD133-stained cells or ( H - I ) tumorsphere formation assay. Respective images and quantitation were shown. Data from three independent experiments in duplicates were presented as means +- SD. ** p < 0.01, *** p < 0.001. Scale bar = 100 mum. ( J ) The Oncomine dataset ""Gaber lung"" was used to analyze USP4 mRNA levels in normal human lung tissues and lung cancers. ( K ) The Oncomine dataset ""Bild lung"" was used to analyze USP4 mRNA levels in stage I or stage II-IV human lung cancers. ( L ) The Oncomine dataset ""Raponi lung"" was used to analyze USP4 mRNA levels in 3 year-alive or 3 year-dead human lung cancer patients.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 Fap1-inhibition decreases abundance of CD133 + CD44 + cells in in a murine xenograft model of colon cancer with or without oxaliplatin Tumors from the mice described above were analyzed for cell population distribution after various treatments. (A) Histograms from flow cytometry demonstrate decreased abundance of CD133 + CD44 + cells after treatment with SLV peptide with or without oxaliplatin. A representative histograms for each cohort is shown. (B) Treatment with SLV peptide decreases relative abundance of CD133 + CD44 + cells in xenograft tumors. Tumors were simultaneously harvested from mice treated with SLV peptide versus VLS control (when control group tumors were >2,000 mm 3 ) and analyzed for CD133 and CD44 expression by flow cytometry. Significant differences indicated by * , ** , or *** . (p2,000 mm 3 ) and analyzed for CD133 and CD44 expression by flow cytometry. Significant differences indicated by * , ** , or *** . (p