Revision 7

#36135

Store at -20C

POU2F3 (E5N2D) XP® Rabbit mAb
CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

Web:

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, IHC-Bond, IHC-P, ChIP, ChIP-seq, C&R

Reactivity:
H

Sensitivity:
Endogenous

MW (kDa):
45-60

Source/Isotype:
Rabbit IgG

UniProt ID:
#Q9UKI9

Entrez-Gene Id:
25833

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
IHC Leica Bond 1:100 - 1:400
Immunohistochemistry (Paraffin) 1:50 - 1:200
Chromatin IP 1:50
Chromatin IP-seq 1:50
CUT&RUN 1:100

Storage

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 #38561.

Specificity/Sensitivity

POU2F3 (E5N2D) XP® Rabbit mAb recognizes endogenous levels of total POU2F3 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu160 of human POU2F3 protein.

Background

POU2F3 (also known as OCT-11 and SKN-1) belongs to the OCT family of octamer motif-binding transcription factors, members of which play varied, diverse roles in animal development. POU2F3 is a lineage-defining transcription factor, responsible for driving differentiation of specialized chemosensory cells in the gastrointestinal and respiratory tracts (1-3). Tuft cells are a subtype of chemosensory cell, defined by POU2F3 expression, and which are hypothesized to be the origin of a distinct, non-neuroendocrine subtype of small cell lung cancer (SCLC) known as tuft cell-like SCLC (4). Consistent with its lineage-defining role in tuft cell development, POU2F3 was identified through a Crispr/Cas9 screen as a master regulator of tuft cell-like SCLC (5).

Background References

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

W: Western Blotting IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq C&R: CUT&RUN

Cross-Reactivity Key

H: Human

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

SignalStain is a registered trademark of Cell Signaling Technology, Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意，否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件，除非书 CST 的合法授书代表以书面形式书独接受， 否书均被拒书，并且无效。

专品专有“专供研究使用”的专专或专似的专专声明， 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的， 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专，且专用于研专用途。将专品用于专断、专防或治专目的， 或专专售（专独或作专专成）或其他商专目的而专专专品，均需要 CST 的专独专可。客专：(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品，或使用专品制造任何商专专品，(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专，或使用专品开专任何与 CST 的专品或服专专争的专品或服专， (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专，(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品 ， (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 7

#36135

POU2F3 (E5N2D) XP® Rabbit mAb

Cell Signaling Technology Logo
Western blot analysis of extracts from NCI-H526, MDA-MB-453, and HCT 116 cells using POU2F3 (E5N2D) XP® Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Expression levels of POU2F3 among cell lines are consistent with expectations based on publicly available bioinformatic databases, confirming specificity of the antibody for POU2F3.
Western Blotting Image 1: POU2F3 (E5N2D) XP<sup>®</sup> Rabbit mAb
Western blot analysis of extracts from 293T cells, mock transfected (lane 1) or transiently transfected with plasmid encoding Myc/DDK-tagged POU2F3 protein (lane 2), using POU2F3 (E5N2D) XP® Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western Blotting Image 2: POU2F3 (E5N2D) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded human small cell lung carcinoma using POU2F3 (E5N2D) XP® Rabbit mAb performed on the Leica® BOND™ Rx.  
Immunohistochemistry Image 1: POU2F3 (E5N2D) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 7

#36135

POU2F3 (E5N2D) XP® Rabbit mAb

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human ovarian clear cell carcinoma using POU2F3 (E5N2D) XP® Rabbit mAb performed on the Leica® BOND™ Rx.  
Immunohistochemistry Image 2: POU2F3 (E5N2D) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded human small cell lung carcinoma using POU2F3 (E5N2D) XP® Rabbit mAb.
Immunohistochemistry Image 1: POU2F3 (E5N2D) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded human large cell neuroendocrine lung carcinoma using POU2F3 (E5N2D) XP® Rabbit mAb.
Immunohistochemistry Image 2: POU2F3 (E5N2D) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 7

#36135

POU2F3 (E5N2D) XP® Rabbit mAb

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human small cell lung carcinoma using POU2F3 (E5N2D) XP® Rabbit mAb.
Immunohistochemistry Image 3: POU2F3 (E5N2D) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded human gastric carcinoma using POU2F3 (E5N2D) XP® Rabbit mAb.
Immunohistochemistry Image 4: POU2F3 (E5N2D) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded normal human skin using POU2F3 (E5N2D) XP® Rabbit mAb.
Immunohistochemistry Image 5: POU2F3 (E5N2D) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 7

#36135

POU2F3 (E5N2D) XP® Rabbit mAb

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded normal human appendix using POU2F3 (E5N2D) XP® Rabbit mAb.
Immunohistochemistry Image 6: POU2F3 (E5N2D) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded normal human parathyroid gland using POU2F3 (E5N2D) XP® Rabbit mAb.
Immunohistochemistry Image 7: POU2F3 (E5N2D) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded normal human kidney using POU2F3 (E5N2D) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right). 
Immunohistochemistry Image 8: POU2F3 (E5N2D) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 7

#36135

POU2F3 (E5N2D) XP® Rabbit mAb

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded NCI-H526 cell pellet (left, positive) or HCT 116 cell pellet (right, negative) using POU2F3 (E5N2D) XP® Rabbit mAb.
Immunohistochemistry Image 9: POU2F3 (E5N2D) XP<sup>®</sup> Rabbit mAb
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCI-H526 cells and POU2F3 (E5N2D) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA library was prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across TRPM5, a known target gene of POU2F3 (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 1: POU2F3 (E5N2D) XP<sup>®</sup> Rabbit mAb
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCI-H526 cells and POU2F3 (E5N2D) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA library was prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 11 (upper), including TRPM5 (lower), a known target gene of POU2F3 (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 2: POU2F3 (E5N2D) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 7

#36135

POU2F3 (E5N2D) XP® Rabbit mAb

Cell Signaling Technology Logo
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCI-H526 cells and either POU2F3 (E5N2D) XP® Rabbit mAb or Normal Rabbit IgG #2729, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR, using human TRPM5 intron 4 primers, human ASCL2 upstream primers, human GFI-B upstream primers, and human IGFBP2 Intron 1 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.
Chromatin Immunoprecipitation Image 3: POU2F3 (E5N2D) XP<sup>®</sup> Rabbit mAb
CUT&RUN was performed with HCT116 cells and POU2F3 (E5N2D) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA library was prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across RPLP1 gene, a known target gene of POU2F3 (see additional figure containing CUT&RUN-qPCR data).
CUT & RUN Image 1: POU2F3 (E5N2D) XP<sup>®</sup> Rabbit mAb
CUT&RUN was performed with HCT116 cells and POU2F3 (E5N2D) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA library was prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 15 (upper), including RPLP1 (lower), a known target gene of POU2F3 (see additional figure containing CUT&RUN-qPCR data).
CUT & RUN Image 2: POU2F3 (E5N2D) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 7

#36135

POU2F3 (E5N2D) XP® Rabbit mAb

Cell Signaling Technology Logo
CUT&RUN was performed with HCT116 cells and either POU2F3 (E5N2D) 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 RPLP1 promoter primers, human LPCAT3 promoter primers, and human IQCF1 intron 2 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 & RUN Image 3: POU2F3 (E5N2D) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.