Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat

Applications

Validated:

Knockout Validated, Western Blot, Immunocytochemistry, Simple Western

Cited:

Western Blot, Flow Cytometry

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 334640
View all RIPK1/RIP1 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human RIPK1/RIP1
Met1-Asn671
Accession # Q13546

Specificity

Detects human, mouse and rat RIPK1/RIP1 in Western blots.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for Human/Mouse/Rat RIPK1/RIP1 Antibody

Detection of Human/Mouse/Rat RIPK1/RIP1 antibody by Western Blot.

Detection of Human/Mouse/Rat RIPK1/RIP1 by Western Blot.

Western blot shows lysates of Raji human Burkitt's lymphoma cell line, Jurkat human acute T cell leukemia cell line, DA3 mouse myeloma cell line, and L6 rat myoblast cell line. PVDF membrane was probed with 0.5 µg/mL of Mouse Anti-Human/Mouse/Rat RIPK1/RIP1 Monoclonal Antibody (Catalog # MAB3585) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for RIPK1/RIP1 at approximately 75 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
RIPK1/RIP1 antibody in MCF-7 Human Cell Line by Immunocytochemistry (ICC).

RIPK1/RIP1 in MCF‑7 Human Cell Line.

RIPK1/RIP1 was detected in immersion fixed MCF-7 human breast cancer cell line using Mouse Anti-Human/Mouse/Rat RIPK1/RIP1 Monoclonal Antibody (Catalog # MAB3585) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Human RIPK1/RIP1 antibody by Simple WesternTM.

Detection of Human RIPK1/RIP1 by Simple WesternTM.

Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line and MCF‑7 human breast cancer cell line, loaded at 0.2 mg/mL. A specific band was detected for RIPK1/RIP1 at approximately 78 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human/Mouse/Rat RIPK1/RIP1 Monoclonal Antibody (Catalog # MAB3585). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.

Western Blot Shows Human RIPK1/RIP1 Antibody Specificity by Using Knockout Cell Line.

Western Blot Shows Human RIPK1/RIP1 Specificity by Using Knockout Cell Line.

Western blot shows lysates of MCF-7 human breast cancer parental cell line and RIPK1/RIP1 knockout MCF-7 cell line (KO). PVDF membrane was probed with 0.5 µg/mL of Mouse Anti-Human/Mouse/Rat RIPK1/RIP1 Monoclonal Antibody (Catalog # MAB3585) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for RIPK1/RIP1 at approximately 75 kDa (as indicated) in the parental MCF-7 cell line, but is not detectable in knockout MCF-7 cell line. GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of RIPK1/RIP1 by Western Blot

Detection of RIPK1/RIP1 by Western Blot

Proteomic analysis identified proteins upregulated in the presence of pro-survival HSP70/BAG1S complex. a U2OS MYC-ER cells expressing ectopic vector, BAG1S, or BAG1 delta S depleted of endogenous BAG1 protein. MYC activity induced for 12 or 24 h with ±100 nM 4-OHT treatment. Lysates analyzed via IB to detect changes in known HSP70 chaperone client proteins GCR, XIAP and RAF1. b Schematic of experimental conditions representing endogenous BAG1 (vector - shLUC), BAG1 knockdown (vector - shBAG1), BAG1S only (BAG1S - shBAG1), or BAG1 delta S only (BAG1 delta S - shBAG1) evaluated for differences in global protein levels. Proteomics analysis outlined with exclusion criteria for significant protein differences between samples. c Efficient knockdown of endogenous BAG1 and rescue of BAG1S and BAG1 delta S shown by IB for samples subjected to proteomics analysis. d Proteomic hits assessed based on schematic of compiled proteins with ≥|1.5| fold change in knockdown compared to control and p ≤ 0.05 across all conditions. Protein expression levels obtained for each sample indexed by specific protein and clustered by UniProt biological process classification. P-values representative of experimental triplicates submitted for proteomic assessment. e Venn diagram showing proteins partially rescued with reintroduction of BAG1S or BAG1 delta S. Increase of ≥10% constitutes a partial rescue. Overlapping proteins with BAG1S or BAG1 delta S indicative of proteins rescued by either ectopic protein. f Verification of proteomics via detection of BAG1S rescued targets SLC7A6 and POLR1D by IB. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30902071), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of RIPK1/RIP1 by Western Blot

Detection of RIPK1/RIP1 by Western Blot

RPE flatmounts are viable for at least one week after culture in vitro. (A) RPE flatmounts were prepared from 2- and 9-month-old wild-type C57BL/6J mice and cultured in complete medium with or without H2O2 for up to one week. Protein lysates were prepared from the RPE flatmounts and subjected to Western blotting analysis. The cell death marker RIP1 was used to evaluate cell viability; vinculin was used as an internal control. As a positive control: RPE flatmounts treated with either 1 mM or 10 mM H2O2 had significant expression of RIP1. Further, the RPE flatmount that was cultured for 7 days without changing the medium (mishandled sample) showed a slight amount of RIP1, whereas the RPE flatmounts cultured with frequent medium changes did not express RIP1, suggesting that the RPE cells obtained even from older mice (9 months) were viable after at least 7 days of culture. (B) RPE flatmounts were obtained from 3-month-old wild-type C57BL/6J mice and cultured in complete medium with or without H2O2 treatment for up to one week. RPE flatmounts were then fixed in 2.5% glutaraldehyde and processed for transmission electron microscopy (TEM). The TEM imaging of the RPE flatmounts cultured for 16 h, 3 days, and 7 days showed that these RPE cells have normal nuclei (yellow arrows) and organelles, such as mitochondria (yellow arrowheads), and abundant microvilli. H2O2-treated RPE flatmounts showed cell death symptoms, including chromatin condensation and fragmentation (red arrow), a disrupted cell membrane, complete loss of microvilli, a translucent cytoplasm, and severely damaged mitochondria (red arrowheads). (C) ZO-1 immunostaining on 2-month-old RPE flatmounts cultured for 3 days showed robust ZO-1 expression, indicating RPE cell integrity. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34769409), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human/Mouse/Rat RIPK1/RIP1 Antibody

Application
Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed MCF-7 human breast cancer cell line

Knockout Validated

RIPK1/RIP1 is specifically detected in MCF‑7 human breast cancer cell line parental cell line but is not detectable in RIPK1/RIP1 knockout MCF‑7 cell line.

Simple Western

10 µg/mL
Sample: Jurkat human acute T cell leukemia cell line and MCF‑7 human breast cancer cell line

Western Blot

0.5 µg/mL
Sample: Raji human Burkitt's lymphoma cell line, Jurkat human acute T cell leukemia cell line, DA3 mouse myeloma cell line, and L6 rat myoblast cell line

Reviewed Applications

Read 2 reviews rated 5 using MAB3585 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: RIPK1/RIP1

Receptor-Interacting Protein 1 (RIP1, also known as RIPK1) is a 671 amino acid (aa) 75 kDa protein that contains an N-terminal protein kinase domain, a C-terminal death domain, and a unique internal region called the intermediate domain. RIP1 is a serine/threonine protein kinase and is constitutively expressed in many tissues. RIP1 interacts with the cytoplasmic death domain of FAS and TNF receptors and is an important element in the signal transduction machinery that mediates apoptosis. RIP1 has been shown to interact with a number of proteins including TRADD, TRAF1, TRAF2, and TRAF3, to form larger signaling complexes. These complexes, in turn, activate specific signaling cascades, such as NF kappa B. RIP1 also interacts through the C-terminal RIP homotypic interaction motif (RHIM) of TRIF in TLR3 dependent activation of NF kappa B.

Long Name

Receptor (TNFRSF)-Interacting Serine-Threonine Kinase 1

Alternate Names

RIP, RIP1

Entrez Gene IDs

8737 (Human); 19766 (Mouse)

Gene Symbol

RIPK1

UniProt

Q13546

Additional RIPK1/RIP1 Products

Product Documents for Human/Mouse/Rat RIPK1/RIP1 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human/Mouse/Rat RIPK1/RIP1 Antibody

For research use only

Citations for Human/Mouse/Rat RIPK1/RIP1 Antibody

Customer Reviews for Human/Mouse/Rat RIPK1/RIP1 Antibody (2)

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  • Human/Mouse/Rat RIPK1/RIP1 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Liver tissue
    Species: Rat
    Verified Customer | Posted 11/08/2023
    Used for rat liver tissue sample at 1:1000 dilution.
    Human/Mouse/Rat RIPK1/RIP1 Antibody MAB3585
  • Human/Mouse/Rat RIPK1/RIP1 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: AML12 (alpha mouse liver 12) cell
    Species: Mouse
    Verified Customer | Posted 05/04/2020
    Aml12 cell line. Cells treated with Nec-1 (RIPK1 inhibitor) for 24 hours. Primary antibody: 1:1000 Secondary antibody: 1: 200000
    Human/Mouse/Rat RIPK1/RIP1 Antibody MAB3585

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