Revision 6

#83413

Store at -20C

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

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

Applications:
W, W-S, IP, IHC-Bond, IHC-P, FC-FP, ChIP

Reactivity:
H

Sensitivity:
Endogenous

MW (kDa):
50

Source/Isotype:
Rabbit IgG

UniProt ID:
#Q02556

Entrez-Gene Id:
3394

Product Usage Information

For optimal ChIP results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 × 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunoprecipitation 1:100
IHC Leica Bond 1:250 - 1:1000
Immunohistochemistry (Paraffin) 1:250 - 1:1000
Flow Cytometry (Fixed/Permeabilized) 1:400 - 1:1600
Chromatin IP 1:50

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 #28852.

Specificity/Sensitivity

IRF-8 (E6J8Q) XP® Rabbit mAb recognizes endogenous levels of total IRF-8 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of human IRF-8 protein.

Background

Interferon regulatory factors (IRFs) comprise a family of transcription factors that function within the Jak/Stat pathway to regulate interferon (IFN) and IFN-inducible gene expression in response to viral infection (1). IRFs play an important role in pathogen defense, autoimmunity, lymphocyte development, cell growth, and susceptibility to transformation. The IRF family includes nine members: IRF-1, IRF-2, IRF-9/ISGF3γ, IRF-3, IRF-4 (Pip/LSIRF/ICSAT), IRF-5, IRF-6, IRF-7, and IRF-8/ICSBP. All IRF proteins share homology in their amino-terminal DNA-binding domains. IRF family members regulate transcription through interactions with proteins that share similar DNA-binding motifs, such as IFN-stimulated response elements (ISRE), IFN consensus sequences (ICS), and IFN regulatory elements (IRF-E) (2).

IRF-8/ICSBP is expressed predominately in hematopoietic cells and is further increased upon treatment with interferon (3,4). IRF-8 can function as a transcription repressor of ICS-containing promoters (4). Expression of IRF-8 can lead to the downregulation of the anti-apoptotic protein Bcl-2 (5). Originally described as being induced by IFN-γ, IRF-8 expression is also elevated by IRF-α as well as IL-12 in NK and T cells (6). IRF-8 deficient mice have enhanced susceptibility to various pathogens and impaired production of interferons, as well as deregulated hematopoiesis that resembles chronic myelogenous leukemia (7,8). IRF-8 also regulates bone metabolism by suppressing osteoclast formation (9).

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 W-S: Simple Western™ IP: Immunoprecipitation IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) FC-FP: Flow Cytometry (Fixed/Permeabilized) ChIP: Chromatin IP

Cross-Reactivity Key

H: Human

Trademarks and Patents

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

Alexa Fluor is a registered trademark of Life Technologies Corporation.

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 6

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using IRF-8 (E6J8Q) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 1: IRF-8 (E6J8Q) XP<sup>®</sup> Rabbit mAb
Simple Western™ analysis of lysates (0.1mg/mL) from Daudi cells using IRF-8 (E6J8Q) XP® Rabbit mAb #83413. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12 – 230 kDa separation module.
Western Blotting Image 1: IRF-8 (E6J8Q) XP<sup>®</sup> Rabbit mAb
Immunoprecipitation of IRF-8 protein from THP-1 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is IRF-8 (E6J8Q) XP® Rabbit mAb. Western blot analysis was performed using IRF-8 (E6J8Q) XP® Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
Immunoprecipitation Image 1: IRF-8 (E6J8Q) 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 6

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human B-cell non-Hodgkin lymphoma using IRF-8 (E6J8Q) XP® Rabbit mAb performed on the Leica® BOND™ Rx.
Immunohistochemistry Image 1: IRF-8 (E6J8Q) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded human T-cell lymphoma using IRF-8 (E6J8Q) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Immunohistochemistry Image 1: IRF-8 (E6J8Q) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded THP-1 cell pellet (left, positive) or Jurkat cell pellet (right, negative) using IRF-8 (E6J8Q) XP® Rabbit mAb.
Immunohistochemistry Image 2: IRF-8 (E6J8Q) 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 6

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using IRF-8 (E6J8Q) XP® Rabbit mAb.
Immunohistochemistry Image 3: IRF-8 (E6J8Q) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded normal human spleen using IRF-8 (E6J8Q) XP® Rabbit mAb.
Immunohistochemistry Image 4: IRF-8 (E6J8Q) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded human gastric adenocarcinoma using IRF-8 (E6J8Q) XP® Rabbit mAb.
Immunohistochemistry Image 5: IRF-8 (E6J8Q) 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 6

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded normal human tonsil using IRF-8 (E6J8Q) XP® Rabbit mAb.
Immunohistochemistry Image 6: IRF-8 (E6J8Q) XP<sup>®</sup> Rabbit mAb
Flow cytometric analysis of HH cells (blue, negative) and THP-1 cells (green, positive) using IRF-8 (E6J8Q) XP® Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor 488® Conjugate) #4412 was used as a secondary antibody.
Flow Cytometry Image 1: IRF-8 (E6J8Q) XP<sup>®</sup> Rabbit mAb
Chromatin immunoprecipitations were performed with cross-linked chromatin from untreated THP-1 cells and either IRF-8 (E6J8Q) 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 SimpleChIP® Human MS4A7 Promoter Primers #9013, hGPR18 promoter primers, 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.
Chromatin Immunoprecipitation Image 1: IRF-8 (E6J8Q) 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.