PRODUCT CODE: ER1902-67

RIP3 Rabbit Polyclonal Antibody (ER1902-67)

Applications

  • WB

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Rat

  • Mouse

Western blot analysis of RIP3 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ER1902-67, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: human lung carcinoma tissue lysate<br />
Lane 2: human placenta tissue lysate
  • Western blot analysis of RIP3 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ER1902-67, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: human lung carcinoma tissue lysate<br />
Lane 2: human placenta tissue lysate
  • Immunohistochemical analysis of paraffin-embedded rat kidney tissue using anti-RIP3 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1902-67, 1/50) for 30 minutes 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 RIP3 was done on SW620 cells. The cells were fixed, permeabilized and stained with the primary antibody (ER1902-67, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of RIP3 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ER1902-67, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: human lung carcinoma tissue lysate
Lane 2: human placenta tissue lysate

Applications

  • WB

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Rat

  • Mouse

SPECIFICATIONS

Product Type

Rabbit polyclonal primary

Product Name

RIP3 Rabbit Polyclonal Antibody (ER1902-67)

Immunogen

Synthetic peptide within human rip3 aa 380-420.

Host

Rabbit

Positive Control

Human lung carcinoma tissue lysate, human placenta tissue lysate, rat kidney tissue, SW620.

Conjugation

Unconjugated

Clonality

Polyclonal

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Peptide affinity purified.

MOLECULAR WEIGHT

57 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB:1:500-1:2,000

  • IHC-P:1:50-1:200

  • FC:1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

RIP3

SYNONYMS

Receptor interacting protein 3 antibody; Receptor interacting serine threonine kinase 3 antibody; Receptor interacting serine/threonine protein kinase 3 antibody; Receptor-interacting protein 3 antibody; Receptor-interacting serine/threonine-protein kinase 3 antibody; RIP 3 antibody; RIP like protein kinase 3 antibody; RIP-3 antibody; RIP-like protein kinase 3 antibody; RIPK 3 antibody; RIPK3 antibody; RIPK3_HUMAN antibody

SEQUENCE SIMILARITIES

Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family.

TISSUE SPECIFICITY

Highly expressed in the pancreas. Detected at lower levels in heart, placenta, lung and kidney. Isoform 3 is significantly increased in colon and lung cancers.

POST-TRANSLATIONAL MODIFICATION

RIPK1 and RIPK3 undergo reciprocal auto- and trans-phosphorylation. Phosphorylation of Ser-199 plays a role in the necroptotic function of RIPK3. Phosphorylation at Ser-227 is required for binding MLKL. Phosphorylation at Thr-182 is important for its kinase activity, interaction with PELI1 and PELI1-mediated 'Lys-48'-linked polyubiquitination and for its ability to mediate TNF-induced necroptosis.; Polyubiquitinated with 'Lys-48' and 'Lys-63'-linked chains by BIRC2/c-IAP1 and BIRC3/c-IAP2, leading to activation of NF-kappa-B. Polyubiquitinated with 'Lys-48'-linked chains by PELI1 leading to its subsequent proteasome-dependent degradation. Ubiquitinated by STUB1 leading to its subsequent proteasome-dependent degradation.

SUBCELLULAR LOCATION

Cytoplasm. Membrane. Mitochondrion.

FUNCTION

Receptor-interacting serine/threonine-protein kinase 3 is an enzyme that in humans is encoded by the RIPK3 gene. The product of this gene is a member of the receptor-interacting protein (RIP) family of serine/threonine protein kinases, and contains a C-terminal domain unique from other RIP family members. The encoded protein is predominantly localized to the cytoplasm, and can undergo nucleocytoplasmic shuttling dependent on novel nuclear localization and export signals. Essential for necroptosis, a programmed cell death process in response to death-inducing TNF-alpha family members. Upon induction of necrosis, RIPK3 interacts with, and phosphorylates RIPK1 and MLKL to form a necrosis-inducing complex. RIPK3 binds to and enhances the activity of three metabolic enzymes: GLUL, GLUD1, and PYGL. These metabolic enzymes may eventually stimulate the tricarboxylic acid cycle and oxidative phosphorylation, which could result in enhanced ROS production.