- 44-711G - Invitrogen Antibodies RELA antibody

Submitted references Mechanical Study of Jian-Gan-Xiao-Zhi Decoction on Nonalcoholic Fatty Liver Disease Based on Integrated Network Pharmacology and Untargeted Metabolomics.

Cao YJ, Li HZ, Zhao J, Sun YM, Jin XW, Lv SQ, Luo JY, Fang XX, Wen WB, Liao JB
Evidence-based complementary and alternative medicine : eCAM 2022;2022:2264394

TNF antagonist sensitizes synovial fibroblasts to ferroptotic cell death in collagen-induced arthritis mouse models.

Wu J, Feng Z, Chen L, Li Y, Bian H, Geng J, Zheng ZH, Fu X, Pei Z, Qin Y, Yang L, Zhao Y, Wang K, Chen R, He Q, Nan G, Jiang X, Chen ZN, Zhu P
Nature communications 2022 Feb 3;13(1):676

Curcumin and α/β-Adrenergic Antagonists Cotreatment Reverse Liver Cirrhosis in Hamsters: Participation of Nrf-2 and NF-κB.

Macías-Pérez JR, Vázquez-López BJ, Muñoz-Ortega MH, Aldaba-Muruato LR, Martínez-Hernández SL, Sánchez-Alemán E, Ventura-Juárez J
Journal of immunology research 2019;2019:3019794

The In Vitro Effect of Acidic-Pepsin on Nuclear Factor KappaB Activation and Its Related Oncogenic Effect on Normal Human Hypopharyngeal Cells.

Sasaki CT, Toman J, Vageli D
PloS one 2016;11(12):e0168269

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot using NF-kappa-B S529 phosphospecific antibody (Product # 44-711G). Jurkat cells were unstimulated (1) or stimulated with PMA and Ca2+ (2-5). Lysates were incubated with anti-NF-kappa-B S529 after prior incubation with: no peptide (1, 2), the non-phosphopeptide (3), a generic [pS] peptide (4), or the phosphopeptide immunogen (5). Data shows antibody phospho-site specificity, and induction of phospho signal after PMA and Ca2+ ionophore treatment.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot using NF-kappa-B S529 phosphospecific antibody (Product # 44-711G). Jurkat cells were unstimulated (1) or stimulated with PMA and Ca2+ (2-5). Lysates were incubated with anti-NF-kappa-B S529 after prior incubation with: no peptide (1, 2), the non-phosphopeptide (3), a generic [pS] peptide (4), or the phosphopeptide immunogen (5). Data shows antibody phospho-site specificity, and induction of phospho signal after PMA and Ca2+ ionophore treatment.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Phospho-NFkB (pS529) showing staining in the nucleus and cytoplasm of paraffin-embedded human spleen tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Phospho-NFkB (pS529) polyclonal antibody (Product # 44-711G) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Phospho-NFkB (pS529) showing staining in the nucleus and cytoplasm of paraffin-embedded mouse spleen tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Phospho-NFkB (pS529) polyclonal antibody (Product # 44-711G) diluted in 3% BSA-PBS at a dilution of 1:50 overnight at 4ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of NFkB [pS529] was done on Jurkat cells treated with TNF alpha (50ng/mL, 20 minutes). Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with NFkB [pS529] Rabbit Polyclonal Antibody (44711G, red histogram) or with rabbit isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Goat Anti-Rabbit Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 1 Effect of pepsin in p-NF-kappaB nuclear localization, in human hypopharyngeal keratinocytes (HHK) and human hypopharyngeal primary cells (HHPC). Immunofluorescence (IF) staining of phospho-NF-kappaB (p-p65 S529) is demonstrated in (A) HHK and (B) HHPC treated by 0.1 mg/ml pepsin at different pH (acid, pH 4.0; weakly acidic, pH 5.0 and neutral, pH 7.0) and corresponding controls. Acidic-pepsin treated HHK and HHPC demonstrates mainly cytoplasmic localization of p-NF-kappaB. In contrast, neutral-pepsin treated HHK and weakly acidic-pepsin treated HHPC results in an intense nuclear localization of p-NF-kappaB.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 2 Varying pepsin concentrations at different pH do not affect the nuclear localization of p-NF-kappaB, in human hypopharyngeal primary cells (HHPC). Immunofluorescence (IF) staining of phospho-NF-kappaB (p-p65 S529) is demonstrated in HHPC treated by 0.01, 0.5 and 01 mg/ml of pepsin at different pH (acid, pH 4.0; weakly acidic, pH 5.0 and neutral, pH 7.0) and corresponding controls.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 3 Effect of pepsin in NF-kappaB activation and bcl-2 overexpression, in human hypopharyngeal keratinocytes (HHK) and human hypopharyngeal primary cells (HHPC). Western blot analysis for NF-kappaB (p65), p-NF-kappaB (p-p65 S529), p-IKB-alpha (Ser32/36) and bcl-2 protein levels demonstrates that acidic-pepsin does not induce NF-kappaB activation or bcl-2 overexpression in treated (A) HHK and (B) HHPC. Columns of the graph correspond to (a) total and nuclear p-NF-kappaB (p-p65) levels, (b) pepsin-induced nuclear p-NF-kappaB (p65 S529) translocation values (p-p65/p65 nuclear/total ratios), (c) cytoplasmic p-IKB-alpha and (d) cytoplasmic/nuclear bcl-2 protein levels (ONE-WAY ANOVA; Kruskal-Wallis; GraphPad Prism 6.0). Mean +-SD of three independent experiments.

44-711G

Antibody data