Invitrogen

RIP1 Recombinant Polyclonal Antibody

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated. View Details

View all (29) RIP1 antibodies

Datasheet

Questions & Answers

Datasheet

Questions & Answers

Product Image Gallery

RIP1 Antibody in Western Blot (WB) — Western blot analysis was performed on Membrane enriched extracts (30 µg lysate) of Jurkat (Lane 1), COLO 205 (Lane 2), SW480 (Lane 3), HT-29 (Lane 4), HCT 116 (Lane 5), MDA-MB-231 (Lane 6), MCF-7 (Lane 7), Hep G2 (Lane 8), A549 (Lane 9) and HeLa (Lane 10). The blots were probed with Anti-RIPK1 Recombinant Rabbit Polyclonal Antibody (Product # 711689, 2.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 76 kDa band corresponding to RIPK1 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0322BOX), 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® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

RIP1 Antibody in Immunocytochemistry (ICC) — Knockdown of RIPK1 was achieved by transfecting HT29 cells with RIPK1 specific siRNA (Silencer® select Cat # s16651 ,s16653). Immunofluorescence analysis was performed on HT29 cells (untransfected, panel a,d), transfected with non-specific scrambled siRNA (panels b,e) and transfected with RIPK1 specific siRNA (panel c,f) Cells were fixed, permeabilized, and labelled with RIPK1 Recombinant Rabbit Polyclonal Antibody (Product # 711689, 5 µg/mL), followed by Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Nuclei (blue) were stained using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938), and Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (red) staining. Loss of signal was observed upon siRNA mediated knockdown (panel c,f) confirming specificity of the antibody to RIPK1 (green). The images were captured at 60X magnification

RIP1 Antibody in Knockdown — Detection and subcellular localization of the target protein by genetic knock down demonstrates antibody specificity. Immunofluorescence analysis of RIPK1 using Anti-RIPK1 Recombinant Rabbit Polyclonal Antibody (Product # 711689) detects RIPK1 in cytoplasm of untransfected and scrambled RNA transfected HT29 cells, where as no signal was observed in RIPK1 specific siRNA transfected cells. {KD}

RIP1 Antibody in Relative expression — Antibody specificity was demonstrated by relative expression. Immunofluorescence analysis of RIPK1 using Anti-RIPK1 Recombinant Rabbit Polyclonal Antibody (Product # 711689) detects RIPK1 in cytoplasm of COLO-205 cells whereas no signal was observed in RIPK1 negative SW480 cells. {RE}

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

2.5 µg/mL

Immunocytochemistry (ICC)

5 µg/mL

Immunofluorescence (IF)

5 µg/mL

Product Specifications

Species Reactivity

Human

Host/Isotype

Rabbit / IgG

Class

Recombinant Polyclonal

Type

Antibody

Immunogen

Protein corresponding to human RIPK1 [aa1-aa294]

Conjugate

Unconjugated

Form

Liquid

Concentration

0.5 mg/mL

Purification

Protein A

Storage buffer

PBS, pH 7.4

Contains

0.09% sodium azide

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RRID

AB_2716903

Product Specific Information

This antibody is predicted to react with Monkey, Horse, Cat

Recombinant rabbit polyclonal antibodies are unique offerings from Thermo Fisher Scientific. They are comprised of a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds - the sensitivity of polyclonal antibodies with the specificity of monoclonal antibodies - all delivered with the consistency only found in a recombinant antibody. While functionally the same as a polyclonal antibody - recognizing multiple epitope sites on the target and producing higher detection sensitivity for low abundance targets - a recombinant rabbit polyclonal antibody has a known mixture of light and heavy chains. The exact population can be produced in every lot, circumventing the biological variability typically associated with polyclonal antibody production.

Target Information

Serine-threonine kinase which transduces inflammatory and cell-death signals (programmed necrosis) following death receptors ligation, activation of pathogen recognition receptors (PRRs), and DNA damage. Upon activation of TNFR1 by the TNF-alpha family cytokines, TRADD and TRAF2 are recruited to the receptor. Phosphorylates DAB2IP at 'Ser-728' in a TNF-alpha-dependent manner, and thereby activates the MAP3K5-JNK apoptotic cascade. Ubiquitination by TRAF2 via 'Lys-63'-link chains acts as a critical enhancer of communication with downstream signal transducers in the mitogen-activated protein kinase pathway and the NF-kappa-B pathway, which in turn mediate downstream events including the activation of genes encoding inflammatory molecules. Polyubiquitinated protein binds to IKBKG/NEMO, the regulatory subunit of the IKK complex, a critical event for NF-kappa-B activation. Interaction with other cellular RHIM-containing adapters initiates gene activation and cell death. RIPK1 and RIPK3 association, in particular, forms a necrosis-inducing complex.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

References (0)

Have you cited this product in a publication?

Let us know so we can reference it here.

Cite this product

Bioinformatics

Protein Aliases: Cell death protein RIP; FLJ39204; OTTHUMP00000015955; receptor (TNFRSF)-interacting serine-threonine kinase 1; Receptor-interacting protein 1; receptor-interacting protein kinase 1; Receptor-interacting serine/threonine-protein kinase 1; RIP-1; RIP1 (CT); serine/threonine-protein kinase RIP

Gene Aliases: RIP; RIP-1; RIP1; RIPK1

UniProt ID: (Human) Q13546

Entrez Gene ID: (Human) 8737

Molecular Function: kinase non-receptor serine/threonine protein kinase non-receptor tyrosine protein kinase protein kinase transferase