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T-bet/TBX21 (D6N8B) XP ® Rabbit mAb
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Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly465 of human T-bet/TBX21 protein.
Product Usage Information
For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Immunoprecipitation | 1:100 |
| Immunohistochemistry (Paraffin) | 1:1600 |
| Immunofluorescence (Immunocytochemistry) | 1:400 |
| Flow Cytometry (Fixed/Permeabilized) | 1:100 - 1:400 |
| Chromatin IP | 1:50 |
| Chromatin IP-seq | 1:50 |
| CUT&RUN | 1:50 |
| CUT&Tag | 1:50 |
Specificity/Sensitivity
Species Reactivity:
Human
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
For a carrier free (BSA and azide free) version of this product see product #27112.
参考图片
CUT&Tag was performed with NK-92 cells and T-bet/TBX21 (D6N8B) XP® Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figures show binding across chromosome 12 (upper), including IFNG (lower), a known target gene of T-bet/TBX21 (see additional figure containing ChIP-qPCR data).
Western blot analysis of extracts from NK-92 and K-562 cells using T-bet/TBX21 (D6N8B) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using T-bet/TBX21 (D6N8B) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded MJ [G11] cell pellet (left, positive) or K-562 cell pellet (right, negative) using T-bet/TBX21 (D6N8B) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded Non-Hodgkin B-cell lymphoma using T-bet/TBX21 (D6N8B) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human tonsil using T-bet/TBX21 (D6N8B) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Immunofluorescent analysis of NK-92 (positive, left) and K-562 (negative, right) cells using T-bet/TBX21 XP® (D6N8B) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Flow cytometric analysis of human PBMCs using T-bet/TBX21 (D6N8B) XP® Rabbit mAb co-stained with either CD56, CD3 or CD19. CD56+ NK cells and a subset of CD3+ T cells are both distinctly positive for T-bet/TBX21 whereas CD19+ B cells are negative. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody for the T-Bet/TBX21 primary.
Chromatin immunoprecipitations were performed with cross-linked chromatin from NK-92 cells and T-bet/TBX21 (D6N8B) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across CCL4, a known target gene of T-bet/TBX21 (see additional figure containing ChIP-qPCR data).
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 NK-92 cells and T-bet/TBX21 (D6N8B) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across chromosome 17 (upper), including CCL4 (lower), a known target gene of T-bet/TBX21 (see additional figure containing ChIP-qPCR data).
Chromatin immunoprecipitations were performed with cross-linked chromatin from NK-92 cells and either T-bet/TBX21 (D6N8B) XP® Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human CCL4 promoter primers, SimpleChIP® Human IFN-γ Promoter Primers #13051, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
CUT&RUN was performed with NK-92 cells and T-bet/TBX21 (D6N8B) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA library was prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figure shows binding across CKS1B gene, a known target gene of T-bet (see additional figure containing CUT&RUN-qPCR data).
CUT&RUN was performed with NK-92 cells and T-bet/TBX21 (D6N8B) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 1 (upper), including CKS1B gene (lower), a known target gene of T-bet (see additional figure containing CUT&RUN-qPCR data).
CUT&RUN was performed with NK-92 cells and T-bet/TBX21 (D6N8B) XP® Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using human GTPbP8 intron 1 primers, human CKS1B exon 1 primers, and human KC6 upstream primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
CUT&Tag was performed with NK-92 cells and T-bet/TBX21 (D6N8B) XP(R) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figure shows binding across IFNG, a known target gene of T-bet/TBX21 (see additional figure containing ChIP-qPCR data).