PA5-16758

antibody from Invitrogen Antibodies

Targeting: RELA NFKB3, p65

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Data presented on provider website)

Immunohistochemistry (Data presented on provider website)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-16758

Provider

Invitrogen Antibodies

Product name

Anti-NFkB p65 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

PA5-16758 targets NFkappaB/p65 in IHC (P) and WB applications and shows reactivity with Human, Rabbit, and Rat samples. The PA5-16758 immunogen is a synthetic peptide corresponding to internal domain of human NFkappaB p65.

Reactivity

Human, Rat

Host

Rabbit

Isotype

IgG

Vial size

500 µL

Storage

-20° C, Avoid Freeze/Thaw Cycles

References

Submitted references

Pretreatment with mineralocorticoid receptor blocker reduces intestinal injury induced by ischemia and reperfusion: involvement of inhibition of inflammatory response, oxidative stress, nuclear factor κB, and inducible nitric oxide synthase.

Ozacmak HS, Ozacmak VH, Barut F, Araslı M, Ucan BH
The Journal of surgical research 2014 Oct;191(2):350-61

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NF-κB and COX-2 expression in nonmalignant endometrial lesions and cancer.

Faloppa CC, Baiocchi G, Cunha IW, Fregnani JH, Osorio CA, Fukazawa EM, Kumagai LY, Badiglian-Filho L, Pinto GL, Soares FA
American journal of clinical pathology 2014 Feb;141(2):196-203

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Mechanisms of the protective effects of curcumin against indomethacin-induced gastric ulcer in rats.

Morsy MA, El-Moselhy MA
Pharmacology 2013;91(5-6):267-74

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot of NFkappaB/p65 using NFkappaB/p65 Polyclonal Antibody (Product # PA5-16758) on LNCaP Cells.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed using whole cell extracts (30 µg lysate) of NIH/3T3 (Lane 1), MCF7 (Lane 2), PC-12 (Lane 3), K-562 (Lane 4) and SH-SY5Y (Lane 5). The blots were probed with Anti-NFkappaB Rabbit polyclonal Antibody (Product # PA5-16758, 2-4 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 65 kDa band corresponding to NFkappaB was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody using iBind™ Flex Western Starter Kit (Product # SLF2000S). Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of NFkappaB / p65 was performed using HeLa cells. 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 NFkappaB Rabbit Polyclonal Antibody (Product # PA5-16758, red histogram) or with rabbit isotype control (pink histogram) at 3-5 µg/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 (Product # 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.