Antibodies, Proteins, Kits and Reagents for Life Science

361 843 ₸ Product size

100 µl 361 843 ₸ 10 µl 147 417 ₸

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Anti-NFkB p105 / p50 antibody [E381] (ab32360)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [E381] to NFkB p105 / p50
Suitable for: WB, IHC-P
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-NFkB p105 / p50 antibody [E381]
See all NFkB p105 / p50 primary antibodies
Description

Rabbit monoclonal [E381] to NFkB p105 / p50
Host species

Rabbit
Specificity

This antibody will detect both forms: p50 and p105.
Tested Applications & Species

Application Species
IHC-P
Human

WB
Rat

Human

See all applications and species data
Immunogen

Synthetic peptide within NFkB p105/ p50 aa 300-400. The exact sequence is proprietary.
Positive control
- WB: HeLa, MCF-7 and PC-12 cell lysates. IHC-P: human prostate carcinoma and bladder carcinoma tissues.
General notes
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 production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Application	Species
IHC-P	Human
WB	Rat Human

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

E381
Isotype

IgG
Research areas
- Cell Biology
- Apoptosis
- Intracellular
- NFkB
- p50
- Cancer
- Cell Death
- Apoptosis
- Apoptosis Markers
- NFkB
- p50

Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)

All lanes : Anti-NFkB p105 / p50 antibody [E381] (ab32360)

Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : NFkB p105 / p50 knockout HAP1 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Jurkat cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 50 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab32360 observed at 120, 50 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab32360 was shown to specifically react with NFkB p105 / p50 when NFkB p105 / p50 knockout samples were used. Wild-type and NFkB p105 / p50 knockout samples were subjected to SDS-PAGE. ab32360 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10 000 respectively and 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 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFkB p105 / p50 antibody [E381] (ab32360)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human bladder carcinoma tissue labelling NFkB p105 / p50 with purified ab32360 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)

All lanes : Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : NFKB1 knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 50 kDa
Observed band size: 105 kDa
why is the actual band size different from the predicted?

Lanes 1- 2: Merged signal (red and green). Green - ab32360 observed at 105 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab32360 was shown to react with NFkB p105 / p50 in wild-type HeLa cells in western blot. The band observed in knockout cell line ab264823 (knockout cell lysate ab257003) lane below 105kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and NFKB1 knockout HeLa 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. ab32360 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.
Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)

Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/10000 dilution (purified) + HeLa cell lysate at 20 µg

Secondary
Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 50 kDa
Observed band size: 105,50 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)

Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/50000 dilution (purified) + MCF-7 cell lysate at 10 µg

Secondary
Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 50 kDa
Observed band size: 105 kDa why is the actual band size different from the predicted?
Additional bands at: 50 kDa (possible non-specific binding)

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)

All lanes : Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/10000 dilution (purified)

Lane 1 : PC-12 cell lysate
Lane 2 : NIH/3T3 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 50 kDa
Observed band size: 105 kDa why is the actual band size different from the predicted?
Additional bands at: 50 kDa (possible non-specific binding)

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)

Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/5000 dilution (unpurified) + HeLa cell lysate

Predicted band size: 50 kDa
Observed band size: 105,50 kDa why is the actual band size different from the predicted?
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFkB p105 / p50 antibody [E381] (ab32360)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma tissue labelling NFkB p105 / p50 with unpurified ab32360 at a dilution of 1/250.
Anti-NFkB p105 / p50 antibody [E381] (ab32360)

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