Buy a SimpleChIP Kit and Try our New SimpleChIP® Universal qPCR Master Mix (88989P) for Free | Get The Code >>
4766
NF-κB Family Member Antibody Sampler Kit

NF-κB Family Member Antibody Sampler Kit #4766

Western Blotting Image 1

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.

Learn more about how we get our images
Western Blotting Image 2

Western blot analysis of extracts from Raji, THP-1 and BaF3 cells using RelB (C1E4) Rabbit mAb.

Learn more about how we get our images
Western Blotting Image 3

Western blot analysis of extracts from K562, Raji and 293 cells, using c-Rel Antibody.

Learn more about how we get our images
Western Blotting Image 4

Western blot analysis of extracts from various cell lines using NF-κB1 p105/p50 (D7H5M) Rabbit mAb.

Learn more about how we get our images
Western Blotting Image 5

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.

Learn more about how we get our images
Western Blotting Image 6

Western blot analysis of extracts from THP-1, L929, C6, and COS cells, using NF-kappaB2 p100 Antibody.

Learn more about how we get our images
Western Blotting Image 7

Western blot analysis of extracts from HeLa, and COS cells, using NF-kB2 p100/p52 (18D10) Rabbit mAb.

Learn more about how we get our images
Chromatin IP-seq Image 8

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 from 5 ng enriched ChIP DNA using NEBNext® Ultra™ II DNA Library Prep Kit for Illumina®, and sequenced on the Illumina NextSeq. 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 data sheet.

Learn more about how we get our images
Flow Cytometry Image 9

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).

Learn more about how we get our images
Western Blotting Image 10

Western blot analysis of extracts from various cell lines using NF-κB p65 (D14E12) XP® Rabbit mAb.

Learn more about how we get our images
Western Blotting Image 11

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.

Learn more about how we get our images
Western Blotting Image 12

Western blot analysis of extracts from various cell lines using NF-κB p65 (L8F6) Mouse mAb.

Learn more about how we get our images
IHC-P (paraffin) Image 13

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using c-Rel Antibody in the presence of control peptide (left) or antigen-specific peptide (right).

Learn more about how we get our images
Western Blotting Image 14

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.

Learn more about how we get our images
IHC-P (paraffin) Image 15

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using NF-kappaB2 p100/p52 (18D10) Rabbit mAb.

Learn more about how we get our images
Chromatin IP Image 16

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.

Learn more about how we get our images
IHC-P (paraffin) Image 17

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)).

Learn more about how we get our images
IHC-P (paraffin) Image 18

Immunohistochemical analysis of human chronic cholecystitis tissue using NF-κB p65 (L8F6) Mouse mAb.

Learn more about how we get our images
IHC-P (paraffin) Image 19

Immunohistochemical analysis of paraffin-embedded Hodgkin's lymphoma, showing nuclear and cytoplasmic localization using c-Rel Antibody.

Learn more about how we get our images
Chromatin IP Image 20

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.

Learn more about how we get our images
IHC-P (paraffin) Image 21

Immunohistochemical analysis of paraffin-embedded human osteosarcoma, using NF-kappaB2 p100/p52 (18D10) Rabbit mAb.

Learn more about how we get our images
IHC-P (paraffin) Image 22

Immunohistochemical analysis of paraffin-embedded human chronic cholecystitis using NF-κB p65 (D14E12) XP® Rabbit mAb.

Learn more about how we get our images
IHC-P (paraffin) Image 23

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.

Learn more about how we get our images
IHC-P (paraffin) Image 24

Immunohistochemical analysis of paraffin-embedded human skin, using c-Rel Antibody.

Learn more about how we get our images
Flow Cytometry Image 25

Flow cytometric analysis of HeLa cells using NF-kappaB2 p100/p52 (18D10) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

Learn more about how we get our images
IHC-P (paraffin) Image 26

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.

Learn more about how we get our images
Flow Cytometry Image 27

Flow cytometric analysis of K562 cells using c-Rel Antibody (blue) compared to a nonspecific negative control antibody (red).

Learn more about how we get our images
IF-IC Image 28

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).

Learn more about how we get our images
Flow Cytometry Image 29

Flow cytometric analysis of HeLa cells using NF-κB p65 (L8F6) Mouse mAb (blue) compared to a nonspecific negative control antibody (red).

Learn more about how we get our images
IF-IC Image 30

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or TNFα-treated (#2169, 20 ng/ml for 20 min, right), using c-Rel Antibody (green). Actin filaments have been labeled with DY-554 phalloidin (red).

Learn more about how we get our images
IF-IC Image 31

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).

Learn more about how we get our images
Chromatin IP Image 32

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.

Learn more about how we get our images
Product Includes Quantity Applications Reactivity MW(kDa) Isotype
NF-κB p65 (L8F6) Mouse mAb 6956 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
  • ChIP
H M R Hm Mk Mi B Dg Pg 65 Mouse IgG2b
RelB (C1E4) Rabbit mAb 4922 20 µl
  • WB
  • IP
H M R Mk 70 Rabbit IgG
c-Rel Antibody 4727 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H Mk 78 Rabbit 
NF-κB1 p105/p50 (D7H5M) Rabbit mAb 12540 20 µl
  • WB
  • IP
  • ChIP
H M 50 Active form. 120 Precursor Rabbit IgG
NF-κB1 p105 Antibody 4717 20 µl
  • WB
  • IP
H M R Mk Mi B Pg 120 Rabbit 
NF-κB2 p100/p52 Antibody 4882 20 µl
  • WB
  • IP
H M R Mk 52 (mature). 120 (precursor). Rabbit 
NF-κB2 p100/p52 (18D10) Rabbit mAb 3017 20 µl
  • WB
  • IHC
  • F
H Mk 52 active form. 120 precursor. Rabbit IgG
NF-κB p65 (D14E12) XP® Rabbit mAb 8242 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
  • ChIP
H M R Hm Mk Dg 65 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl
  • WB
Horse 

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).

Each antibody in this kit recognizes endogenous levels of its target protein regardless of post-translational modification state such as phosphorylation or acetylation. The NF-κB1 p105/p50 (D7H5M) Rabbit mAb #12540 detects both the precursor protein p105 and its cleavage product p50, while the NF-κB1 p105 Antibody #4717 only detects p105 and will not cross-react with p50. Both the NF-κB2 p100/p52 Antibody #4882 and the NF-κB2 p100/p52 (18D10) Rabbit mAb (Human Specific) #3017 will cross-react with the precursor protein p100 and its cleavage product p52.

Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to amino acid residues near the carboxy terminus of human c-Rel, at the carboxy terminus of human NF-κB1 p105 and near the amino terminus of human NF-κB2 p100/p52. Antibodies are purified by Protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to amino acid residues near the carboxy terminus of human NF-κB p65, surrounding Ser424 of human RelB, surrounding Gly415 of human NF-κB p105/p50 protein, surrounding Glu498 of human NF-κB p65/RelA protein, and near the amino terminus of human NF-κB2 p100/p52.

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.
Entrez-Gene Id
4790 , 5970 , 4791 , 5966 , 5971
Swiss-Prot Acc.
P19838 , Q04206 , Q00653 , Q04864 , Q01201
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

Upstream / Downstream

pathwayImage

Explore pathways related to this product.