Tollip Antibody (503315)

R&D Systems | Catalog # MAB4678

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human, Rat

Cited:

Fish - Danio rerio (Zebrafish)

Applications

Validated:

Western Blot

Cited:

Immunohistochemistry, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2A Clone # 503315
View all Tollip Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human Tollip
Met1-Pro274
Accession # Q9H0E2

Specificity

Detects human and rat Tollip in Western blots.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2A

Scientific Data Images for Tollip Antibody (503315)

Detection of Human/Rat Tollip antibody by Western Blot.

Detection of Human/Rat Tollip by Western Blot.

Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line and NRK rat normal kidney cell line. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human/Rat Tollip Monoclonal Antibody (Catalog # MAB4678) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Tollip at approximately 28 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
Detection of Tollip by Immunohistochemistry

Detection of Tollip by Immunohistochemistry

Knockout of the tollip gene has no effect on the fish morphology. (A) Wild‐type (tollip+/+) and mutant (tollip−/−) embryos at 14 h, and 1 and 5 days post fertilization (hpf, dpf) show normal development and morphology. Scale bar 500 μm. Right image, wild‐type (tollip+/+) and mutant (tollip−/−) male adults at 14 mpf are without visible body morphological defects. Scale bar 1 cm. (B) Morphology of wild‐type tollip+/+ and mutant tollip−/− embryos injected with 2 ng of translation‐blocking morpholino targeting tollip (MO), observed at 48 hpf. Scale bar 1 mm. (C–H) Histology and quantitative analysis of hematoxylin and eosin stained sections of testes (C), spleen (D), liver (E, F), and anterior intestine (G, H) with specific cells and structures marked, including spermatozoa (Sm), spermatogonia (Sg), spermatocytes (Sp), red pulp (RP), white pulp (WP), blood vessels (BV), liver macrophages (arrows in E), and goblet cells (gc). For liver analyses, areas of hepatocytes and their nuclei are quantified in F. The height of intestinal folds is quantified in H. Scale bar 50 µm (C, G), 100 µm (D, E). Bars represent the means ± SEM (n = 3, each encompassing 40–60 (liver) or 25 (intestine) measurements from 5 selected areas) t‐test, ns.—nonsignificant. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35506194), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Tollip by Western Blot

Detection of Tollip by Western Blot

CRISPR/Cas9‐based genome editing allows for the generation of a Tollip‐deficient zebrafish line. (A) Schemes of the zebrafish tollip transcript variants 1 and 2 (v1 and v2), based on the Ensembl database, showing exons (E), translated sequences (gray), and UTR regions (white). (B) Schematic illustration of the structure of Tollip protein isoforms with the C2 and CUE domains indicated. (C) Partial DNA sequence of the target site within exon 2 of the tollip gene in wild‐type tollip+/+ fish (left) and homozygous tollip−/− knockout fish (right). Deletion of eight nucleotides observed in the mutant line is shadowed in dark gray. There is an additional nucleotide change flanking the deletion (double peak marked R in the chromatogram, corresponding to A or G, with a predicted amino acid change D to G in the truncated protein product), indicating mosaicism of the generated line. (D) Schematic illustration of the predicted structure of Tollip protein isoforms synthesized from the mutated tollip gene. (E) Western blot of the 5 dpf protein lysates from the wild‐type (tollip+/+) line and tollip−/− siblings. Top panel shows Tollip (~ 35 kDa) and a bottom panel shows alpha ‐tubulin (~ 55 kDa) signal. (F) qPCR analysis of the expression of tollip transcripts during early zebrafish development (1–5 dpf). Bars represent the means ± SEM from 3–4 independent experiments (encompassing a pool of 10 larvae/condition). Mann–Whitney U test, *P < 0.05, ****P < 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35506194), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Tollip by Immunohistochemistry

Detection of Tollip by Immunohistochemistry

Knockout of the tollip gene has no effect on the fish morphology. (A) Wild‐type (tollip+/+) and mutant (tollip−/−) embryos at 14 h, and 1 and 5 days post fertilization (hpf, dpf) show normal development and morphology. Scale bar 500 μm. Right image, wild‐type (tollip+/+) and mutant (tollip−/−) male adults at 14 mpf are without visible body morphological defects. Scale bar 1 cm. (B) Morphology of wild‐type tollip+/+ and mutant tollip−/− embryos injected with 2 ng of translation‐blocking morpholino targeting tollip (MO), observed at 48 hpf. Scale bar 1 mm. (C–H) Histology and quantitative analysis of hematoxylin and eosin stained sections of testes (C), spleen (D), liver (E, F), and anterior intestine (G, H) with specific cells and structures marked, including spermatozoa (Sm), spermatogonia (Sg), spermatocytes (Sp), red pulp (RP), white pulp (WP), blood vessels (BV), liver macrophages (arrows in E), and goblet cells (gc). For liver analyses, areas of hepatocytes and their nuclei are quantified in F. The height of intestinal folds is quantified in H. Scale bar 50 µm (C, G), 100 µm (D, E). Bars represent the means ± SEM (n = 3, each encompassing 40–60 (liver) or 25 (intestine) measurements from 5 selected areas) t‐test, ns.—nonsignificant. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35506194), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Tollip by Immunohistochemistry

Detection of Tollip by Immunohistochemistry

Knockout of the tollip gene has no effect on the fish morphology. (A) Wild‐type (tollip+/+) and mutant (tollip−/−) embryos at 14 h, and 1 and 5 days post fertilization (hpf, dpf) show normal development and morphology. Scale bar 500 μm. Right image, wild‐type (tollip+/+) and mutant (tollip−/−) male adults at 14 mpf are without visible body morphological defects. Scale bar 1 cm. (B) Morphology of wild‐type tollip+/+ and mutant tollip−/− embryos injected with 2 ng of translation‐blocking morpholino targeting tollip (MO), observed at 48 hpf. Scale bar 1 mm. (C–H) Histology and quantitative analysis of hematoxylin and eosin stained sections of testes (C), spleen (D), liver (E, F), and anterior intestine (G, H) with specific cells and structures marked, including spermatozoa (Sm), spermatogonia (Sg), spermatocytes (Sp), red pulp (RP), white pulp (WP), blood vessels (BV), liver macrophages (arrows in E), and goblet cells (gc). For liver analyses, areas of hepatocytes and their nuclei are quantified in F. The height of intestinal folds is quantified in H. Scale bar 50 µm (C, G), 100 µm (D, E). Bars represent the means ± SEM (n = 3, each encompassing 40–60 (liver) or 25 (intestine) measurements from 5 selected areas) t‐test, ns.—nonsignificant. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35506194), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Tollip by Immunohistochemistry

Detection of Tollip by Immunohistochemistry

Knockout of the tollip gene has no effect on the fish morphology. (A) Wild‐type (tollip+/+) and mutant (tollip−/−) embryos at 14 h, and 1 and 5 days post fertilization (hpf, dpf) show normal development and morphology. Scale bar 500 μm. Right image, wild‐type (tollip+/+) and mutant (tollip−/−) male adults at 14 mpf are without visible body morphological defects. Scale bar 1 cm. (B) Morphology of wild‐type tollip+/+ and mutant tollip−/− embryos injected with 2 ng of translation‐blocking morpholino targeting tollip (MO), observed at 48 hpf. Scale bar 1 mm. (C–H) Histology and quantitative analysis of hematoxylin and eosin stained sections of testes (C), spleen (D), liver (E, F), and anterior intestine (G, H) with specific cells and structures marked, including spermatozoa (Sm), spermatogonia (Sg), spermatocytes (Sp), red pulp (RP), white pulp (WP), blood vessels (BV), liver macrophages (arrows in E), and goblet cells (gc). For liver analyses, areas of hepatocytes and their nuclei are quantified in F. The height of intestinal folds is quantified in H. Scale bar 50 µm (C, G), 100 µm (D, E). Bars represent the means ± SEM (n = 3, each encompassing 40–60 (liver) or 25 (intestine) measurements from 5 selected areas) t‐test, ns.—nonsignificant. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35506194), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Tollip Antibody (503315)

Application
Recommended Usage

Western Blot

1 µg/mL
Sample: HepG2 human hepatocellular carcinoma cell line and NRK rat normal kidney cells

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: Tollip

Tollip (Toll-interacting protein) is a 28 kDa, cytoplasmic protein that is involved in IL-1 and Toll receptor signaling. It is classified as an adaptor protein that performs multiple functions. It binds both IRAK and Toll receptors, inhibiting constitutive IL-1 and Toll receptor signaling. It also binds to ubiquinated IL-1 RI, directing it to the endosomal degradation compartment. Human Tollip is 274 amino acids(aa) in length. It contains a C2 Ca++-binding region (aa 54‑154) and a CUE domain (aa 229‑270) that binds ubiquitin. One potential splice form exists that shows a deletion of aa 6‑33. Human Tollip is 92% identical to mouse Tollip.

Long Name

Toll Interacting Protein

Alternate Names

IL-1RAcPIP

Entrez Gene IDs

54472 (Human); 54473 (Mouse); 361677 (Rat)

Gene Symbol

TOLLIP

UniProt

Q9H0E2 (Human)

Additional Tollip Products

Product Documents for Tollip Antibody (503315)

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Product Specific Notices for Tollip Antibody (503315)

For research use only

Citations for Tollip Antibody (503315)

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