Mouse FIZZ1/RELM alpha Antibody (226033)

R&D Systems | Catalog # MAB1523

R&D Systems

Key Product Details

Species Reactivity

Validated:

Mouse

Cited:

Mouse, Transgenic Mouse

Applications

Validated:

Western Blot, Immunocytochemistry

Cited:

Immunohistochemistry, Flow Cytometry, Fluorescence Microscopy

Label

Unconjugated

Antibody Source

Monoclonal Rat IgG2A Clone # 226033
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Product Specifications

Immunogen

E. coli-derived recombinant mouse FIZZ1/RELM alpha
Asp24-Ser111
Accession # Q9EP95

Specificity

Detects mouse FIZZ1/RELM alpha in direct ELISAs and Western blots. In direct ELISAs and Western blots, no cross-reactivity with recombinant mouse (rm) RELM beta or rmResistin is observed.

Clonality

Monoclonal

Host

Rat

Isotype

IgG2A

Scientific Data Images for Mouse FIZZ1/RELM alpha Antibody (226033)

FIZZ1/RELMa antibody in BV-2 Mouse Cell Line by Immunocytochemistry (ICC).

FIZZ1/RELM alpha in BV-2 Mouse Cell Line.

FIZZ1/RELMa was detected in immersion fixed BV-2 mouse microglia cell line using Rat Anti-Mouse FIZZ1/RELMa Monoclonal Antibody (Catalog # MAB1523) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (orange; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of FIZZ1/RELM alpha by Western Blot

Detection of FIZZ1/RELM alpha by Western Blot

OTUD6A deubiquitinates hResistin in the endoplasmic reticulum to divert it from proteasomal degradation to secretion. A‐B) Co‐immunoprecipitation of endogenous OTUD6A and mRELM alpha in HDM‐CAM (A) and HDM‐AAM (B) lung tissues. C) Co‐immunoprecipitation of exogenous OTUD6A and hResistin in BEAS‐2B cells expressed Flag‐OTUD6A and HA‐hResistin. D) Immunofluorescence of hResistin and OTUD6A in BEAS‐2B cells transfected with Flag‐OTUD6A and HA‐hResistin. Scale bars: 20 µm. E) CHX chase assay for hResistin stability (n = 3). F) BEAS‐2B cells were stimulated with DMSO, MG132 (10 µm), Epoxomicin (2.5 µm), BTZ (5 µm), CQ (25 µm), 3‐MA (10 mm), Bafilomycin A1 (1 µm), or Concanamycin A (0.5 µm) for 6 h. Western blot analysis of hResistin in BEAS‐2B cells. G) HA‐hResistin, Flag‐OTUD6A, and Myc‐Ub were transfected into BEAS‐2B cells and then subjected to 10 µm MG132 for 6 h. Ubiquitinated hResistin was detected. H) HA‐hResistin, Flag‐OTUD6A, Myc‐WT Ub, Myc‐K48 Ub, and Myc‐K63 Ub were transfected into BEAS‐2B cells and then subjected to 10 µm MG132 for 6 h. Ubiquitinated hResistin was detected. I) BEAS‐2B cells were stimulated with DMSO, MG132 (10 µm), Epoxomicin (2.5 µm), or BTZ (5 µm) for 6 h or transfected with Flag‐OTUD6A for 24 h, ELISA analysis of hResistin levels in cell supernatant (n = 5). J) BEAS‐2B cells transfected with Myc‐K48 Ub or Myc‐K63 Ub for 24 h, ELISA analysis of hResistin levels in cell supernatant (n = 5). K) Immunofluorescence of Resistin and Calnexin in BEAS‐2B cells treated with MG132 (10 µm) or not for 6 h. Scale bars: 50 µm. L) Immunofluorescence of Resistin and OTUD6A in BEAS‐2B cells treated with MG132 (10 µm) or not for 6 h. Scale bars: 50 µm. Data are presented as mean ± SEM. P values determined by one‐way ANOVA (*p < 0.05, **p < 0.01, ***p < 0.001). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/41560298), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of FIZZ1/RELM alpha by Western Blot

Detection of FIZZ1/RELM alpha by Western Blot

OTUD6A deubiquitinates hResistin in the endoplasmic reticulum to divert it from proteasomal degradation to secretion. A‐B) Co‐immunoprecipitation of endogenous OTUD6A and mRELM alpha in HDM‐CAM (A) and HDM‐AAM (B) lung tissues. C) Co‐immunoprecipitation of exogenous OTUD6A and hResistin in BEAS‐2B cells expressed Flag‐OTUD6A and HA‐hResistin. D) Immunofluorescence of hResistin and OTUD6A in BEAS‐2B cells transfected with Flag‐OTUD6A and HA‐hResistin. Scale bars: 20 µm. E) CHX chase assay for hResistin stability (n = 3). F) BEAS‐2B cells were stimulated with DMSO, MG132 (10 µm), Epoxomicin (2.5 µm), BTZ (5 µm), CQ (25 µm), 3‐MA (10 mm), Bafilomycin A1 (1 µm), or Concanamycin A (0.5 µm) for 6 h. Western blot analysis of hResistin in BEAS‐2B cells. G) HA‐hResistin, Flag‐OTUD6A, and Myc‐Ub were transfected into BEAS‐2B cells and then subjected to 10 µm MG132 for 6 h. Ubiquitinated hResistin was detected. H) HA‐hResistin, Flag‐OTUD6A, Myc‐WT Ub, Myc‐K48 Ub, and Myc‐K63 Ub were transfected into BEAS‐2B cells and then subjected to 10 µm MG132 for 6 h. Ubiquitinated hResistin was detected. I) BEAS‐2B cells were stimulated with DMSO, MG132 (10 µm), Epoxomicin (2.5 µm), or BTZ (5 µm) for 6 h or transfected with Flag‐OTUD6A for 24 h, ELISA analysis of hResistin levels in cell supernatant (n = 5). J) BEAS‐2B cells transfected with Myc‐K48 Ub or Myc‐K63 Ub for 24 h, ELISA analysis of hResistin levels in cell supernatant (n = 5). K) Immunofluorescence of Resistin and Calnexin in BEAS‐2B cells treated with MG132 (10 µm) or not for 6 h. Scale bars: 50 µm. L) Immunofluorescence of Resistin and OTUD6A in BEAS‐2B cells treated with MG132 (10 µm) or not for 6 h. Scale bars: 50 µm. Data are presented as mean ± SEM. P values determined by one‐way ANOVA (*p < 0.05, **p < 0.01, ***p < 0.001). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/41560298), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of FIZZ1/RELM alpha by Western Blot

Detection of FIZZ1/RELM alpha by Western Blot

hResistin/mRELM alpha is a potential substrate of OTUD6A. A) Multi‐omics identification of OTUD6A substrates. B‐C) Western blot of mRELM alpha expression in HDM‐CAM (B) and HDM‐AAM (C) lung tissues. D‐E) IHC staining of mRELM alpha in HDM‐CAM (D) and HDM‐AAM (E) lung tissues. Scale bars: 100 µm. F‐G) Correlation analysis of mRELM alpha and OTUD6A IHC staining in HDM‐CAM (F) and HDM‐AAM (G). Two random fields per mouse. H) Western blot analysis of hResistin in BEAS‐2B cells transfected with Flag‐OTUD6A and control vector for 24 h. I) Western blot analysis of hResistin in HBEpiC cells transfected with Flag‐OTUD6A and control vector for 24 h. J) Western blot analysis of hResistin in BEAS‐2B cells transfected with siOTUD6A for 24 h with or without HDM (100 µg/mL) treatment for 6 h. K) Western blot analysis of hResistin in HBEpiC cells transfected with siOTUD6A for 24 h with or without HDM (100 µg/mL) treatment for 6 h. Data are presented as mean ± SEM. P values determined by two‐tailed unpaired t‐test or one‐way ANOVA (*p < 0.05, **p < 0.01, ***p < 0.001). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/41560298), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Mouse FIZZ1/RELM alpha Antibody (226033)

Application
Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed BV-2 mouse microglia cell line

Western Blot

1 µg/mL
Sample: Recombinant Mouse FIZZ1/RELM alpha

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.


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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: FIZZ1/RELM alpha

RELM alpha, also known as ‘found in inflammatory zone 1’ (FIZZ1) belongs to the RELM/FIZZ family of cysteine rich secretory proteins. RELM alpha is most abundantly expressed in white adipose tissue but not in preadipocytes or 3T3-L1 adipocytes. RELM alpha expression is also detected in mammary tissue, heart, lung, and tongue.

Long Name

Found in Inflammatory Zone 1/Resistin-like Molecule alpha

Alternate Names

RELM alpha, RETNLA, RETNLB

Entrez Gene IDs

84666 (Human); 57262 (Mouse)

Gene Symbol

RETNLB

UniProt

Additional FIZZ1/RELM alpha Products

Product Documents for Mouse FIZZ1/RELM alpha Antibody (226033)

Certificate of Analysis

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Product Specific Notices for Mouse FIZZ1/RELM alpha Antibody (226033)

For research use only

Citations for Mouse FIZZ1/RELM alpha Antibody (226033)

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Protocols

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