AB109440

Anti-NFkB p100/NFKB2 antibody [EPR4686]

Name: Anti-NFkB p100/NFKB2 antibody [EPR4686] (ab109440) | Abcam
Brand: Abcam Limited
SKU: AB109440
Price: 395 USD
Availability: InStock
Rating: 5 (2 reviews)

RabMAb
Recombinant
KO Validated
What is this?

(2 Reviews)

(7 Publications)

Rabbit Recombinant Monoclonal NFkB p100/NFKB2 antibody. Suitable for WB, ICC/IF and reacts with Human samples. Cited in 7 publications.

View Alternative Names

LYT10, NFKB2, Nuclear factor NF-kappa-B p100 subunit, DNA-binding factor KBF2, H2TF1, Lymphocyte translocation chromosome 10 protein, Nuclear factor of kappa light polypeptide gene enhancer in B-cells 2, Oncogene Lyt-10, Lyt10

7 Images

Immunocytochemistry/ Immunofluorescence - Anti-NFkB p100/NFKB2 antibody [EPR4686] (AB109440)

ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-NFkB p100/NFKB2 antibody [EPR4686] (AB109440)

ab109440 staining NFkB p100/p52 in wild-type HAP1 cells (top panel) and NFkB p100/p52 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min) permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab109440 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

Lab

Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] (AB109440)

Lanes 1- 2 : Merged signal (red and green). Green - ab109440 observed at 120 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab109440 was shown to react with NFkB p100/NFKB2 in wild-type HepG2 cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab262323 (CRISPR/Cas9 edited cell lysate ab257247) lane below 97kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HepG2 and NFKB2 CRISPR/Cas9 edited HepG2 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109440 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] (ab109440) at 1/1000 dilution

Lane 1:

Wild-type HepG2 cell lysate at 20 µg

Lane 2:

Western blot - Human NFKB2 (NFkB p100/NFKB2) knockout Hep G2 cell lysate (<a href='/en-us/products/cell-lysates/human-nfkb2-nfkb-p100-nfkb2-knockout-hep-g2-cell-lysate-ab257247'>ab257247</a>) at 20 µg

Secondary

Lanes 1 - 2:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Lanes 1 - 2:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Predicted band size: 30 kDa,97 kDa

Observed band size: 120 kDa,30 kDa,37 kDa

false

Lab

Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] (AB109440)

Lanes 1- 2 : Merged signal (red and green). Green - ab109440 observed at 120 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab109440 was shown to react with NFkB p100/NFKB2 in wild-type HCT116 cells in western blot. The band observed in knockout cell line ab266883 (knockout cell lysate ab257245) lane below 97kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HCT116 and NFKB2 knockout HCT116 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109440 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] (ab109440) at 1/1000 dilution

Lane 1:

Wild-type HCT116 cell lysate at 20 µg

Lane 2:

NFKB2 knockout HCT116 cell lysate at 20 µg

Predicted band size: 97 kDa

Observed band size: 120 kDa

false

Lab

Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] (AB109440)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)

Lane 2 : NFκB p100 knockout HAP1 cell lysate (20 μg)

Lane 3 : Jurkat cell lysate (20 μg)

Lane 4 : HeLa cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab109440 observed at 100 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab109440 was shown to specifically react with NFκB p100 when NFκB p100 knockout samples were used. Wild-type and NFκB p100 knockout samples were subjected to SDS-PAGE. ab109440 and ab109440 (loading control to GAPDH) were diluted 1/10000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] (ab109440)

Predicted band size: 97 kDa

false

Lab

Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] (AB109440)

Lanes 1- 2 : Merged signal (red and green). Green - ab109440 observed at 120 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab109440 was shown to react with NFkB p100/NFKB2 in wild-type HCT116 cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab266883 (CRISPR/Cas9 edited cell lysate ab257245) lane below 97kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HCT116 and NFKB2 CRISPR/Cas9 edited HCT116 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109440 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] (ab109440) at 1/1000 dilution

Lane 1:

Wild-type HCT116 cell lysate at 20 µg

Lane 2:

NFKB2 CRISPR/Cas9 edited HCT116 cell lysate at 20 µg

Lane 2:

Western blot - Human NFKB2 (NFkB p100/NFKB2) knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-nfkb2-nfkb-p100-nfkb2-knockout-hct116-cell-line-ab266883'>ab266883</a>)

Predicted band size: 97 kDa

Observed band size: 120 kDa

false

Unknown

Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] (AB109440)

All lanes:

Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] (ab109440) at 1/10000 dilution

Lane 1:

Jurkat cell lysate at 10 µg

Lane 2:

HeLa cell lysate at 10 µg

Lane 3:

ECV-304 cell lysate at 10 µg

Lane 4:

MCF7 cell lysate at 10 µg

Predicted band size: 97 kDa

Observed band size: 110 kDa

false

Lab

Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] (AB109440)

Lanes 1- 2 : Merged signal (red and green). Green - ab109440 observed at 120 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab109440 was shown to react with NFkB p100/NFKB2 in wild-type HepG2 cells in western blot. The band observed in knockout cell line ab262323 (knockout cell lysate ab257247) lane below 97kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HepG2 and NFKB2 knockout HepG2 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109440 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] (ab109440) at 1/1000 dilution

Lane 1:

Wild-type HepG2 cell lysate at 20 µg

Lane 2:

NFKB2 knockout HeLa cell lysate at 20 µg

Predicted band size: 97 kDa

Observed band size: 120 kDa

false

Carrier free

Anti-NFkB p100/NFKB2 antibody [EPR4686] - BSA and Azide free

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR4686

Isotype

IgG

Carrier free

Reacts with

Human

Applications

ICC/IF, WB

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "FlowCyt" : {"fullname" : "Flow Cytometry", "shortname":"Flow Cyt"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/10000 - 1/50000", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/250", "ICCIF-species-notes": "" }, "Mouse": { "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "", "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "ICCIF-species-checked": "predicted", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "" } } }

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Product details

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Species reactivity
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species.
Please contact us for more information.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Preservative: 0.05% Sodium azide Constituents: 50% Tissue culture supernatant, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 0.1% BSA

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Storage information

Stable for 12 months at -20°C

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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Target data

NF-kappa-B is a pleiotropic transcription factor present in almost all cell types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NF-kappa-B is a homo- or heterodimeric complex formed by the Rel-like domain-containing proteins RELA/p65, RELB, NFKB1/p105, NFKB1/p50, REL and NFKB2/p52. The dimers bind at kappa-B sites in the DNA of their target genes and the individual dimers have distinct preferences for different kappa-B sites that they can bind with distinguishable affinity and specificity. Different dimer combinations act as transcriptional activators or repressors, respectively. NF-kappa-B is controlled by various mechanisms of post-translational modification and subcellular compartmentalization as well as by interactions with other cofactors or corepressors. NF-kappa-B complexes are held in the cytoplasm in an inactive state complexed with members of the NF-kappa-B inhibitor (I-kappa-B) family. In a conventional activation pathway, I-kappa-B is phosphorylated by I-kappa-B kinases (IKKs) in response to different activators, subsequently degraded thus liberating the active NF-kappa-B complex which translocates to the nucleus. In a non-canonical activation pathway, the MAP3K14-activated CHUK/IKKA homodimer phosphorylates NFKB2/p100 associated with RelB, inducing its proteolytic processing to NFKB2/p52 and the formation of NF-kappa-B RelB-p52 complexes. The NF-kappa-B heterodimeric RelB-p52 complex is a transcriptional activator. The NF-kappa-B p52-p52 homodimer is a transcriptional repressor. NFKB2 appears to have dual functions such as cytoplasmic retention of attached NF-kappa-B proteins by p100 and generation of p52 by a cotranslational processing. The proteasome-mediated process ensures the production of both p52 and p100 and preserves their independent function. p52 binds to the kappa-B consensus sequence 5'-GGRNNYYCC-3', located in the enhancer region of genes involved in immune response and acute phase reactions. p52 and p100 are respectively the minor and major form; the processing of p100 being relatively poor. Isoform p49 is a subunit of the NF-kappa-B protein complex, which stimulates the HIV enhancer in synergy with p65. In concert with RELB, regulates the circadian clock by repressing the transcriptional activator activity of the CLOCK-BMAL1 heterodimer.

See full target information NFKB2

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Publications (7)

Recent publications for all applications. Explore the full list and refine your search

Frontiers in physiology 13:934899 PubMed35991177

2022

I-κB kinase-ε deficiency improves doxorubicin-induced dilated cardiomyopathy by inhibiting the NF-κB pathway.

Applications

Unspecified application

Species

Unspecified reactive species

Yafeng Liu,Yueyue Xu,Yiwei Yao,Yide Cao,Ganyi Chen,Yuchen Cai,Wen Chen,Xin Chen,Zhibing Qiu

Experimental & molecular medicine 53:1623-1635 PubMed34702956

2021

Upregulating sirtuin 6 ameliorates glycolysis, EMT and distant metastasis of pancreatic adenocarcinoma with krüppel-like factor 10 deficiency.

Applications

Unspecified application

Species

Unspecified reactive species

Yi-Chih Tsai,Su-Liang Chen,Shu-Ling Peng,Ya-Li Tsai,Zuong-Ming Chang,Vincent Hung-Shu Chang,Hui-Ju Ch'ang

American journal of human genetics 108:1026-1039 PubMed34004138

2021

Altered replication stress response due to CARD14 mutations promotes recombination-induced revertant mosaicism.

Applications

Unspecified application

Species

Unspecified reactive species

Toshinari Miyauchi,Shotaro Suzuki,Masae Takeda,Jin Teng Peh,Masayuki Aiba,Ken Natsuga,Yasuyuki Fujita,Takuya Takeichi,Taiko Sakamoto,Masashi Akiyama,Hiroshi Shimizu,Toshifumi Nomura

Oxidative medicine and cellular longevity 2021:6429197 PubMed33628362

2021

I-B Kinase- Deficiency Attenuates the Development of Angiotensin II-Induced Myocardial Hypertrophy in Mice.

Applications

Unspecified application

Species

Unspecified reactive species

Yide Cao,Liangpeng Li,Yafeng Liu,Ganyi Chen,Zhonghao Tao,Rui Wang,Wen Chen

Molecular & cellular proteomics : MCP 18:1307-1319 PubMed30992313

2019

High-throughput Exploration of the Network Dependent on AKT1 in Mouse Ovarian Granulosa Cells.

Applications

Unspecified application

Species

Unspecified reactive species

Maëva Elzaiat,Laetitia Herman,Bérangère Legois,Thibaut Léger,Anne-Laure Todeschini,Reiner A Veitia

Oncogenesis 8:13 PubMed30783098

2019

An FBXW7-ZEB2 axis links EMT and tumour microenvironment to promote colorectal cancer stem cells and chemoresistance.

Applications

Unspecified application

Species

Unspecified reactive species

Ningning Li,Roya Babaei-Jadidi,Federica Lorenzi,Bradley Spencer-Dene,Philip Clarke,Enric Domingo,Eugene Tulchinsky,Robert G J Vries,David Kerr,Yihang Pan,Yulong He,David O Bates,Ian Tomlinson,Hans Clevers,Abdolrahman S Nateri

Nature methods 15:909-912 PubMed30377371

2018

A high-throughput pipeline for validation of antibodies.

Applications

Unspecified application

Species

Unspecified reactive species

Krzysztof Sikorski,Adi Mehta,Marit Inngjerdingen,Flourina Thakor,Simon Kling,Tomas Kalina,Tuula A Nyman,Maria Ekman Stensland,Wei Zhou,Gustavo A de Souza,Lars Holden,Jan Stuchly,Markus Templin,Fridtjof Lund-Johansen

View all publications

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Anti-NFkB p100/NFKB2 antibody [EPR4686]

Immunocytochemistry/ Immunofluorescence - Anti-NFkB p100/NFKB2 antibody [EPR4686] (AB109440)

Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] (AB109440)

Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] (AB109440)

Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] (AB109440)

Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] (AB109440)

Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] (AB109440)

Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] (AB109440)

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Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

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Reactivity data

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Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Storage information

Product protocols

Target data

Publications (7)

Product promise