N4BP1 Recombinant Rabbit Monoclonal Antibody [JE63-75]
Catalog# HA722576
N4BP1 Recombinant Rabbit Monoclonal Antibody [JE63-75]
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WB
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IF-Cell
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IHC-P
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FC
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Mouse
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Rat
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unconjugated
Overview
Product Name
N4BP1 Recombinant Rabbit Monoclonal Antibody [JE63-75]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within mouse N4BP1 aa 251-500 / 893.
Species Reactivity
Mouse, Rat
Validated Applications
WB, IF-Cell, IHC-P, FC
Molecular Weight
Predicted band size: 99 kDa
Positive Control
F9 cell lysate, C2C12 cell lysate, Neuro-2a cell lysate, PC-12 cell lysate, C2C12, mouse testis tissue, rat testis tissue.
Conjugation
unconjugated
Clone Number
JE63-75
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:1,000
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IF-Cell
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1:100
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IHC-P
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1:1,000
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FC
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1:1,000
Target
Function
Potent suppressor of cytokine production that acts as a regulator of innate immune signaling and inflammation. Acts as a key negative regulator of select cytokine and chemokine responses elicited by TRIF-independent Toll-like receptors (TLRs), thereby limiting inflammatory cytokine responses to minor insults. In response to more threatening pathogens, cleaved by CASP8 downstream of TLR3 or TLR4, leading to its inactivation, thereby allowing production of inflammatory cytokines (By similarity). Acts as a restriction factor against some viruses, such as HIV-1: restricts HIV-1 replication by binding to HIV-1 mRNAs and mediating their degradation via its ribonuclease activity. Also acts as an inhibitor of the E3 ubiquitin-protein ligase ITCH: acts by interacting with the second WW domain of ITCH, leading to compete with ITCH's substrates and impairing ubiquitination of substrates (By similarity).
Background References
1. Ma Z et al. N4BP1 mediates RAM domain-dependent notch signaling turnover during neocortical development. EMBO J. 2023 Nov
2. Wu C et al. N4BP1 regulates keratinocytes development and plays protective role in burn- and adhesive-related skin injury via MMP9. Cell Signal. 2023 Oct
Subcellular Location
Cytoplasm, cytosol, Nucleus, nucleolus, PML body.
Synonyms
FLJ31821 antibody
KIAA0615 antibody
MGC176730 antibody
N4BP1 antibody
N4BP1_HUMAN antibody
NEDD4 binding protein 1 antibody
NEDD4-binding protein 1 antibody
Images
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Western blot analysis of N4BP1 on different lysates with Rabbit anti-N4BP1 antibody (HA722576) at 1/1,000 dilution.
Lane 1: F9 cell lysate
Lane 2: C2C12 cell lysate
Lane 3: Neuro-2a cell lysate
Lane 4: PC-12 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 99 kDa
Observed band size: 110 kDa
Exposure time: 59 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722576) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of C2C12 cells labeling N4BP1 with Rabbit anti-N4BP1 antibody (HA722576) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-N4BP1 antibody (HA722576) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-N4BP1 antibody (HA722576) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722576) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-N4BP1 antibody (HA722576) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722576) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Flow cytometric analysis of C2C12 cells labeling N4BP1.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA722576, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"