Revision 1

#4766Store at -20C

1 Kit

(8 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
NF-κB p65 (L8F6) Mouse mAb 6956 20 µl 65 kDa Mouse IgG2b
RelB (C1E4) Rabbit mAb 4922 20 µl 70 kDa Rabbit IgG
c-Rel (D4Y6M) Rabbit mAb 12707 20 µl 68-78 kDa Rabbit IgG
NF-κB1 p105/p50 (D7H5M) Rabbit mAb 12540 20 µl 50 Active form. 120 Precursor kDa Rabbit IgG
NF-κB1 p105 Antibody 4717 20 µl 120 kDa Rabbit 
NF-κB2 p100/p52 Antibody 4882 20 µl 52 (mature). 120 (precursor). kDa Rabbit 
NF-κB2 p100/p52 (18D10) Rabbit mAb 3017 20 µl 52 active form. 120 precursor. kDa Rabbit IgG
NF-κB p65 (D14E12) XP® Rabbit mAb 8242 20 µl 65 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl Horse 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

This kit contains reagents to examine total protein levels of the five NF-κB/Rel family members: p65/RelA, RelB, c-Rel, NF-κB1 (p105/p50) and NF-κB2 (p100/p52).

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.

Background

Transcription factors of the nuclear factor κB (NF-κB)/Rel family play a pivotal role in inflammatory and immune responses (1,2). There are five family members in mammals: RelA, c-Rel, RelB, NF-κB1 (p105/p50), and NF-κB2 (p100/p52). Both p105 and p100 are proteolytically processed by the proteasome to produce p50 and p52, respectively. Rel proteins bind p50 and p52 to form dimeric complexes that bind DNA and regulate transcription. In unstimulated cells, NF-κB is sequestered in the cytoplasm by IκB inhibitory proteins (3-5). NF-κB-activating agents can induce the phosphorylation of IκB proteins, targeting them for rapid degradation through the ubiquitin-proteasome pathway and releasing NF-κB to enter the nucleus where it regulates gene expression (6-8). NIK and IKKα (IKK1) regulate the phosphorylation and processing of NF-κB2 (p100) to produce p52, which translocates to the nucleus (9-11).

  1. Baeuerle, P.A. and Henkel, T. (1994) Annu Rev Immunol 12, 141-79.
  2. Baeuerle, P.A. and Baltimore, D. (1996) Cell 87, 13-20.
  3. Haskill, S. et al. (1991) Cell 65, 1281-9.
  4. Thompson, J.E. et al. (1995) Cell 80, 573-82.
  5. Whiteside, S.T. et al. (1997) EMBO J 16, 1413-26.
  6. Traenckner, E.B. et al. (1995) EMBO J 14, 2876-83.
  7. Scherer, D.C. et al. (1995) Proc Natl Acad Sci USA 92, 11259-63.
  8. Chen, Z.J. et al. (1996) Cell 84, 853-62.
  9. Senftleben, U. et al. (2001) Science 293, 1495-9.
  10. Coope, H.J. et al. (2002) EMBO J 21, 5375-85.
  11. Xiao, G. et al. (2001) Mol Cell 7, 401-9.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    Limited Uses

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    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    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 1
    #4766

    NF-κB Family Member Antibody Sampler Kit

    NF-κB Family Member Antibody Sampler Kit: Image 1 Expand Image
    Immunoprecipitation of NF-kB p65 from HeLa cell extracts. Lane 1 is 10% input, lane 2 is precipitated with Mouse (G3A1) mAb IgG1 Isotype Control #5415, and lane 3 is NF-κB p65 (L8F6) Mouse mAb, #6956. Western blot was performed using NF-κB p65 (D14E12) XP® Rabbit mAb, #8242.
    NF-κB Family Member Antibody Sampler Kit: Image 2 Expand Image

    Immunoprecipitation of NF-kB p65 from HeLa cell extracts. Lane 1 is 10% input, lane 2 is precipitated with Mouse (G3A1) mAb IgG1 Isotype Control #5415, and lane 3 is NF-κB p65 (L8F6) Mouse mAb, #6956. Western blot was performed using NF-κB p65 (D14E12) XP® Rabbit mAb, #8242.

    NF-κB Family Member Antibody Sampler Kit: Image 3 Expand Image
    Simple Western™ analysis of lysates (1 mg/mL) from HeLa cells using NF-κB p65 (D14E12) XP® Rabbit mAb #8242. The virtual lane view (left) shows a single 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.
    NF-κB Family Member Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from various cell lines using NF-κB1 p105/p50 (D7H5M) Rabbit mAb.
    NF-κB Family Member Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from various cell lines and tissues using c-Rel (D4Y6M) Rabbit mAb.
    NF-κB Family Member Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of extracts from HeLa, and COS cells, using NF-kB2 p100/p52 (18D10) Rabbit mAb.
    NF-κB Family Member Antibody Sampler Kit: Image 7 Expand Image
    CUT&RUN was performed with HDLM-2 cells and either NF-κB2 p100/p52 (18D10) Rabbit mAb, NF-κB2 p100/p52 (D7A9K) Rabbit mAb #37359, or NF-κB2 p100/p52 (D9S3M) Rabbit mAb #52583, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across BIRC3 gene.
    NF-κB Family Member Antibody Sampler Kit: Image 8 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma using NF-κB2 p100/p52 (18D10) Rabbit mAb.
    NF-κB Family Member Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of extracts from THP-1 cells, differentiated with TPA (#9905, 80 nM for 24 h) and treated with 1 μg/ml LPS for the indicated times, using NF-κB1 p105 Antibody.
    NF-κB Family Member Antibody Sampler Kit: Image 10 Expand Image
    Western blot analysis of extracts from THP-1, L929, C6, and COS cells, using NF-kappaB2 p100 Antibody.
    NF-κB Family Member Antibody Sampler Kit: Image 11 Expand Image
    Western blot analysis of extracts from Raji, THP-1 and BaF3 cells using RelB (C1E4) Rabbit mAb.
    NF-κB Family Member Antibody Sampler Kit: Image 12 Expand Image
    CUT&RUN was performed with HDLM-2 cells and RelB (C1E4) 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 BIRC3 gene.
    NF-κB Family Member Antibody Sampler Kit: Image 13 Expand Image
    Western blot analysis of extracts from control HeLa cells (lane 1) or NF-κB p65 knockout HeLa cells (lane 2) using NF-κB p65 (L8F6) Mouse mAb #6956 (upper) or β-actin (13E5) Rabbit mAb #4970 (lower). The absence of signal in the NF-κB p65 knockout HeLa cells confirms the specificity of the antibody for NF-κB p65.
    NF-κB Family Member Antibody Sampler Kit: Image 14 Expand Image
    Flow cytometric analysis of MCF7 cells using NF-kB p65 (L8F6) Mouse mAb (solid line) or concentration-matched Mouse (G3A1) mAb IgG1 Isotype Control #5415 (dashed lines). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.
    NF-κB Family Member Antibody Sampler Kit: Image 15 Expand Image
    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
    NF-κB Family Member Antibody Sampler Kit: Image 16 Expand Image
    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO* is added and emits light during enzyme catalyzed decomposition.
    NF-κB Family Member Antibody Sampler Kit: Image 17 Expand Image
    Western blot analysis of extracts from various cell lines using NF-κB p65 (D14E12) XP® Rabbit mAb.
    NF-κB Family Member Antibody Sampler Kit: Image 18 Expand Image
    Western blot analysis of extracts from HeLa cells, untreated (-) or treated with Human Tumor Necrosis Factor-α (hTNF-α) #8902 (10 ng/ml) for the indicated times, using NF-κB1 p105/p50 (D7H5M) Rabbit mAb.
    NF-κB Family Member Antibody Sampler Kit: Image 19 Expand Image
    Western blot analysis of extracts from Neuro-2a cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® c-Rel siRNA I (Mouse Specific) #13058 (+) or SignalSilence® c-Rel siRNA II (Mouse Specific) #13170 (+), using c-Rel (D4Y6M) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The c-Rel (D4Y6M) Rabbit mAb confirms silencing of c-Rel expression, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.
    NF-κB Family Member Antibody Sampler Kit: Image 20 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using NF-κB2 p100/p52 (18D10) Rabbit mAb.
    NF-κB Family Member Antibody Sampler Kit: Image 21 Expand Image
    CUT&RUN was performed with HDLM-2 cells and either NF-κB2 p100/p52 (18D10) Rabbit mAb, NF-κB2 p100/p52 (D7A9K) Rabbit mAb #37359, or NF-κB2 p100/p52 (D9S3M) Rabbit mAb #52583, 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 11 (upper), including BIRC3 gene (lower).
    NF-κB Family Member Antibody Sampler Kit: Image 22 Expand Image
    Immunohistochemical analysis of paraffin-embedded HCT 116 cell pellet (left, high-expressing) or MCF7 cell pellet (right, low-expressing) using NF-κB2 p100/p52 (18D10) Rabbit mAb.
    NF-κB Family Member Antibody Sampler Kit: Image 23 Expand Image
    CUT&RUN was performed with HDLM-2 cells and RelB (C1E4) 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 figures show binding across chromosome 11 (upper), including BIRC3 gene (lower).
    NF-κB Family Member Antibody Sampler Kit: Image 24 Expand Image
    Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-) or SignalSilence® NF-κB p65 siRNA I #6261 (+), using NF-κB p65 (L8F6) Mouse mAb (upper) or α-Tubulin (11Η10) Rabbit mAb #2125 (lower). The NF-κB p65 (L8F6) Mouse mAb confirms silencing of NF-κB p65 expression, while the α-Tubulin (11Η10) Rabbit mAb is used as a loading control.
    NF-κB Family Member Antibody Sampler Kit: Image 25 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells treated with Human Tumor Necrosis Factor-α (hTNF-α) #8902 (30 ng/ml) for 1 hour and either NF-κB1 p105/p50 (D7H5M) 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 SimpleChIP® Human IκBα Promoter Primers #5552, human IAP2 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.
    NF-κB Family Member Antibody Sampler Kit: Image 26 Expand Image
    Immunohistochemical analysis of paraffin-embedded human osteosarcoma, using NF-κB2 p100/p52 (18D10) Rabbit mAb.
    NF-κB Family Member Antibody Sampler Kit: Image 27 Expand Image
    CUT&RUN was performed with HDLM-2 cells and either NF-κB2 p100/p52 (18D10) 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 SimpleChIP® Human IκBα Promoter Primers #5552 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.
    NF-κB Family Member Antibody Sampler Kit: Image 28 Expand Image
    CUT&RUN was performed with HDLM-2 cells and either RelB (C1E4) 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 SimpleChIP® Human IκBα Promoter Primers #5552 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.
    NF-κB Family Member Antibody Sampler Kit: Image 29 Expand Image
    Western blot analysis of extracts from various cell lines using NF-κB p65 (L8F6) Mouse mAb.
    NF-κB Family Member Antibody Sampler Kit: Image 30 Expand Image
    Immunohistochemical analysis using NF-κB p65 (D14E12) XP® Rabbit mAb on SignalSlide® NF-κB p65 IHC Controls #12873 (paraffin-embedded HCT116 cells, untreated (left) or treated with hTNF-α #8902 (right)).
    NF-κB Family Member Antibody Sampler Kit: Image 31 Expand Image
    Immunohistochemical analysis of human chronic cholecystitis tissue using NF-κB p65 (L8F6) Mouse mAb.
    NF-κB Family Member Antibody Sampler Kit: Image 32 Expand Image
    Immunohistochemical analysis of paraffin-embedded human chronic cholecystitis using NF-κB p65 (D14E12) XP® Rabbit mAb.
    NF-κB Family Member Antibody Sampler Kit: Image 33 Expand Image
    Immunohistochemical analysis of paraffin-embedded human B-cell non-Hodgkin lymphoma using NF-κB1 p105/p50 (D7H5M) Rabbit mAb.
    NF-κB Family Member Antibody Sampler Kit: Image 34 Expand Image
    Immunohistochemical analysis of paraffin-embedded OVCAR8 cell pellets treated with Human Tumor Necrosis Factor-α (hTNF-α) #8902 (left) or treated with SignalSilence® NF-κB p65 siRNA I #6261 (right), using NF-κB p65 (L8F6) Mouse mAb.
    NF-κB Family Member Antibody Sampler Kit: Image 35 Expand Image
    Confocal immunofluorescent analysis of HT-1080 cells, untreated (left) or treated with hTNF-α #8902 (20 ng/ml, 20 min) (right), using NF-κB p65 (D14E12) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    NF-κB Family Member Antibody Sampler Kit: Image 36 Expand Image
    Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using NF-κB1 p105/p50 (D7H5M) Rabbit mAb.
    NF-κB Family Member Antibody Sampler Kit: Image 37 Expand Image
    Flow cytometric analysis of HeLa cells using NF-κB2 p100/p52 (18D10) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).
    NF-κB Family Member Antibody Sampler Kit: Image 38 Expand Image
    Immunohistochemical analysis of paraffin-embedded HeLa cell pellets, untreated (left) or treated with Human Tumor Necrosis Factor-α (hTNF-α) #8902 (right), using NF-κB p65 (L8F6) Mouse mAb.
    NF-κB Family Member Antibody Sampler Kit: Image 39 Expand Image
    Flow cytometric analysis of HeLa cells using NF-κB p65 (D14E12) XP® Rabbit mAb (blue) compared to concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red).
    NF-κB Family Member Antibody Sampler Kit: Image 40 Expand Image
    Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using NF-κB1 p105/p50 (D7H5M) Rabbit mAb.
    NF-κB Family Member Antibody Sampler Kit: Image 41 Expand Image
    Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with Human Tumor Necrosis Factor-α (hTNF-α) #8902 (20 ng/mL, 20 min; right), using NF-κB p65 (L8F6) Mouse mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    NF-κB Family Member Antibody Sampler Kit: Image 42 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells treated with hTNF-α #8902 (30 ng/ml, 1 hr) and NF-κB p65 (D14E12) XP® Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across IL-8, a known target gene of NFκB (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product datasheet.
    NF-κB Family Member Antibody Sampler Kit: Image 43 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using NF-κB1 p105/p50 (D7H5M) Rabbit mAb.
    NF-κB Family Member Antibody Sampler Kit: Image 44 Expand Image
    Flow cytometric analysis of MCF7 cells using NF-kB p65 (L8F6) Mouse mAb (solid line) compared to a concentration-matched Mouse (G3A1) mAb IgG1 Isotype Control #5415 (dashed lines). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.
    NF-κB Family Member Antibody Sampler Kit: Image 45 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells treated with hTNF-α #8902 (30 ng/ml, 1 hr) and NF-κB p65 (D14E12) XP® Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 4 (upper), including IL-8 (lower), a known target gene of NFκB (see additional figure containing ChIP-qPCR data).
    NF-κB Family Member Antibody Sampler Kit: Image 46 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human thymus using NF-κB1 p105/p50 (D7H5M) Rabbit mAb.
    NF-κB Family Member Antibody Sampler Kit: Image 47 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells treated with Human Tumor Necrosis Factor-α (hTNF-α) #8902 (30 ng/ml, 1 hr) and either NF-κB p65 (L8F6) Mouse 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 SimpleChIP® Human IκBα Promoter Primers #5552, human IL-8 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.
    NF-κB Family Member Antibody Sampler Kit: Image 48 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells treated with hTNF-α #8902 (30 ng/ml, 1 hr) and either NF-κB p65 (D14E12) 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 SimpleChIP® Human IκBα Promoter Primers #5552, human IL-8 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.
    NF-κB Family Member Antibody Sampler Kit: Image 49 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse spleen using NF-κB1 p105/p50 (D7H5M) Rabbit mAb.
    NF-κB Family Member Antibody Sampler Kit: Image 50 Expand Image
    CUT&RUN was performed with HeLa cells treated with hTNF-α #8902 (30 ng/ml, 1 hr) and NF-κB p65 (D14E12) 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 figure shows binding across LAMC2, a known target gene of NF-κB p65 (see additional figure containing CUT&RUN-qPCR data).
    NF-κB Family Member Antibody Sampler Kit: Image 51 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse thymus using NF-κB1 p105/p50 (D7H5M) Rabbit mAb.
    NF-κB Family Member Antibody Sampler Kit: Image 52 Expand Image
    CUT&RUN was performed with HeLa cells treated with hTNF-α #8902 (30 ng/ml, 1 hr) and NF-κB p65 (D14E12) 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 LAMC2 (lower), a known target gene of NF-κB p65 (see additional figure containing CUT&RUN-qPCR data).
    NF-κB Family Member Antibody Sampler Kit: Image 53 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma (left), normal human spleen (middle), or 4T1 syngeneic mammary tumor (right) using NF-κB1 p105/p50 (D7H5M) Rabbit mAb (top) or NF-κB1 p105/p50 Rabbit mAb (bottom). These two antibodies detect unique, non-overlapping epitopes on NF-κB1 p105/p50 protein. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining.
    NF-κB Family Member Antibody Sampler Kit: Image 54 Expand Image
    CUT&RUN was performed with HeLa cells treated with hTNF-α #8902 (30 ng/ml, 1 hr) and either NF-κB p65 (D14E12) 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 LAMC2 upstream primers, and human ITM2A 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.
    NF-κB Family Member Antibody Sampler Kit: Image 55 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human appendix using NF-κB1 p105/p50 (D7H5M) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
    NF-κB Family Member Antibody Sampler Kit: Image 56 Expand Image
    Immunoprecipitation of NF-kB p65 from CHO cell extracts. Lane 1 is 10% input, lane 2 is precipitated with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is NF-κB p65 (D14E12) XP® Rabbit mAb, #8242. Western blot was performed using NF-κB p65 (L8F6) Mouse mAb, #6956.
    NF-κB Family Member Antibody Sampler Kit: Image 57 Expand Image
    Immunohistochemical analysis of paraffin-embedded MEF cell pellet, wild-type (left, positive) or NF-κB1 knockout (right, negative), using NF-κB1 p105/p50 (D7H5M) Rabbit mAb.
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    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.