UNC13D/Munc 13-4 Antibody

Novus Biologicals | Catalog # NB100-41385

Novus Biologicals
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Key Product Details

Validated by

Knockout/Knockdown, Biological Validation

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Predicted:

Bovine (100%), Canine (100%), Porcine (100%), Rat (100%). Backed by our 100% Guarantee.

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Peptide ELISA, Immunocytochemistry/ Immunofluorescence, Knockdown Validated

Cited:

Western Blot, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all UNC13D Primary Antibodies »

Product Specifications

Immunogen

Peptide with sequence C-ETQKHKKDLHPLFD corresponding to internal region according to NP_954712.1.

Reactivity Notes

Predicted cross-reactivity based on sequence identity: Rat, Canine, Porcine, Bovine.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for UNC13D/Munc 13-4 Antibody

Western Blot: UNC13D/Munc 13-4 Antibody [NB100-41385]

Western Blot: UNC13D/Munc 13-4 Antibody [NB100-41385]

Western Blot: UNC13D/Munc 13-4 Antibody [NB100-41385] - RBL-2H3 from Wt or shRNA Munc13-4 lentiviral stable expression whole cell lysate. 50 ug total protein. WB image submitted by a verified customer review.
Immunohistochemistry-Paraffin: UNC13D/Munc 13-4 Antibody [NB100-41385]

Immunohistochemistry-Paraffin: UNC13D/Munc 13-4 Antibody [NB100-41385]

Immunohistochemistry-Paraffin: UNC13D/Munc 13-4 Antibody [NB100-41385] - Staining of paraffin embedded Mouse Lung with antibody at 0.5 ug/mL (wt in A and KO in B).
Western Blot: UNC13D/Munc 13-4 Antibody [NB100-41385]

Western Blot: UNC13D/Munc 13-4 Antibody [NB100-41385]

Western Blot: UNC13D/Munc 13-4 Antibody [NB100-41385] - Staining of Human T-lymphocyte lysate with antibody at 2 ug/mL (35 ug protein in RIPA buffer). Detected by chemiluminescence.
Immunohistochemistry-Paraffin: UNC13D/Munc 13-4 Antibody [NB100-41385]

Immunohistochemistry-Paraffin: UNC13D/Munc 13-4 Antibody [NB100-41385]

Immunohistochemistry-Paraffin: UNC13D/Munc 13-4 Antibody [NB100-41385] - Staining of paraffin embedded Human Thymus. Antibody at 2.5 ug/mL. Steamed antigen retrieval with citrate buffer pH 6, AP-staining.
Knockdown Validated: UNC13D/Munc 13-4 Antibody [NB100-41385]

Western Blot: UNC13D/Munc 13-4 Antibody [NB100-41385]

UNC13D-Munc-13-4-Antibody-Knockdown-Validated-NB100-41385-img0009.jpg
Knockdown Validated: UNC13D/Munc 13-4 Antibody [NB100-41385]

Western Blot: UNC13D/Munc 13-4 Antibody [NB100-41385]

UNC13D-Munc-13-4-Antibody-Knockdown-Validated-NB100-41385-img0010.jpg
UNC13D/Munc 13-4 Antibody

Western Blot: UNC13D/Munc 13-4 Antibody [NB100-41385] -

Western Blot: UNC13D/Munc 13-4 Antibody [NB100-41385] - Munc13-4 KD strongly impairs exosome release. (A) SDS-PAGE Western blot of indicated proteins in MDA-MB-231 cells after stable expression of shRNA for Munc13-4 or Rab27a or a scrambled control (Ctrl). (B) Culture medium from MDA-MB-231 cells either untreated or stimulated w/ 1.25 µM ionomycin for 30 min centrifuged at 1,000 g to remove cellular debris & 10,000 g to remove large extracellular vesicles. (C) The resulting 10,000-g supernatant filtered onto a nitrocellulose membrane & analyzed for CD63, CD9, ALIX, & GM130 content by antibody blotting. (D) Quantification of CD63, CD9, & ALIX blots in C are shown as exosome release as a percentage of total cellular material w/ mean values ± standard error (SE) for n ≥ 3. *, P < 0.05 for comparison w/ corresponding control samples. (E) Panc-1 or A549 cells left untreated or treated w/ TGF beta -1 for 24 h. Indicated proteins detected by SDS-PAGE Western blot. (F) Panc-1 cells left untreated or treated w/ TGF beta -1 for 24 h, & Munc13-4 levels determined by immunofluorescence. TGF beta -1–treated cells exhibited a mesenchymal morphology. Bars, 5 μm. (G) A549 cells stably expressing control shRNA (Ctrl) or Munc13-4 shRNA left untreated (Untr) or treated w/ TGF beta -1 for 24 h, & SDS-PAGE Western blotting for indicated proteins conducted. (H) Culture media supernatants (as in B) from A549 cells that either untreated or stimulated w/ 1.25 µM ionomycin for 30 min filtered onto nitrocellulose membrane & analyzed for CD63 & GM130. (I) Quantification of CD63+ exosome release shown as a percentage of total cellular material w/ mean values ± SE for n = 5. *, P < 0.05; **, P < 0.01 for comparison w/ corresponding control samples. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29930202), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
UNC13D/Munc 13-4 Antibody

Western Blot: UNC13D/Munc 13-4 Antibody [NB100-41385] -

Western Blot: UNC13D/Munc 13-4 Antibody [NB100-41385] - Munc13-4 KD strongly impairs exosome release. (A) SDS-PAGE Western blot of indicated proteins in MDA-MB-231 cells after stable expression of shRNA for Munc13-4 or Rab27a or a scrambled control (Ctrl). (B) Culture medium from MDA-MB-231 cells either untreated or stimulated w/ 1.25 µM ionomycin for 30 min centrifuged at 1,000 g to remove cellular debris & 10,000 g to remove large extracellular vesicles. (C) The resulting 10,000-g supernatant filtered onto a nitrocellulose membrane & analyzed for CD63, CD9, ALIX, & GM130 content by antibody blotting. (D) Quantification of CD63, CD9, & ALIX blots in C are shown as exosome release as a percentage of total cellular material w/ mean values ± standard error (SE) for n ≥ 3. *, P < 0.05 for comparison w/ corresponding control samples. (E) Panc-1 or A549 cells left untreated or treated w/ TGF beta -1 for 24 h. Indicated proteins detected by SDS-PAGE Western blot. (F) Panc-1 cells left untreated or treated w/ TGF beta -1 for 24 h, & Munc13-4 levels determined by immunofluorescence. TGF beta -1–treated cells exhibited a mesenchymal morphology. Bars, 5 μm. (G) A549 cells stably expressing control shRNA (Ctrl) or Munc13-4 shRNA left untreated (Untr) or treated w/ TGF beta -1 for 24 h, & SDS-PAGE Western blotting for indicated proteins conducted. (H) Culture media supernatants (as in B) from A549 cells that either untreated or stimulated w/ 1.25 µM ionomycin for 30 min filtered onto nitrocellulose membrane & analyzed for CD63 & GM130. (I) Quantification of CD63+ exosome release shown as a percentage of total cellular material w/ mean values ± SE for n = 5. *, P < 0.05; **, P < 0.01 for comparison w/ corresponding control samples. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29930202), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
UNC13D/Munc 13-4 Antibody

Western Blot: UNC13D/Munc 13-4 Antibody [NB100-41385] -

Western Blot: UNC13D/Munc 13-4 Antibody [NB100-41385] - Munc13-4 KD strongly impairs exosome release. (A) SDS-PAGE Western blot of indicated proteins in MDA-MB-231 cells after stable expression of shRNA for Munc13-4 or Rab27a or a scrambled control (Ctrl). (B) Culture medium from MDA-MB-231 cells either untreated or stimulated w/ 1.25 µM ionomycin for 30 min centrifuged at 1,000 g to remove cellular debris & 10,000 g to remove large extracellular vesicles. (C) The resulting 10,000-g supernatant filtered onto a nitrocellulose membrane & analyzed for CD63, CD9, ALIX, & GM130 content by antibody blotting. (D) Quantification of CD63, CD9, & ALIX blots in C are shown as exosome release as a percentage of total cellular material w/ mean values ± standard error (SE) for n ≥ 3. *, P < 0.05 for comparison w/ corresponding control samples. (E) Panc-1 or A549 cells left untreated or treated w/ TGF beta -1 for 24 h. Indicated proteins detected by SDS-PAGE Western blot. (F) Panc-1 cells left untreated or treated w/ TGF beta -1 for 24 h, & Munc13-4 levels determined by immunofluorescence. TGF beta -1–treated cells exhibited a mesenchymal morphology. Bars, 5 μm. (G) A549 cells stably expressing control shRNA (Ctrl) or Munc13-4 shRNA left untreated (Untr) or treated w/ TGF beta -1 for 24 h, & SDS-PAGE Western blotting for indicated proteins conducted. (H) Culture media supernatants (as in B) from A549 cells that either untreated or stimulated w/ 1.25 µM ionomycin for 30 min filtered onto nitrocellulose membrane & analyzed for CD63 & GM130. (I) Quantification of CD63+ exosome release shown as a percentage of total cellular material w/ mean values ± SE for n = 5. *, P < 0.05; **, P < 0.01 for comparison w/ corresponding control samples. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29930202), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
UNC13D/Munc 13-4 Antibody

Immunocytochemistry/ Immunofluorescence: UNC13D/Munc 13-4 Antibody [NB100-41385] -

Munc13-4 translocation to membrane is Ca2+ dependent. (A) Live-cell epifluorescence imaging of GFP-Munc13-4 in MDA-MB-231 cells at indicated times after ionomycin stimulation. See Video 1. (B) MDA-MB-231 cells expressing wild-type GFP-Munc13-4, GFP-Munc13-4 C2A*, or GFP-Munc13-4 C2B* either left untreated or stimulated with 1.25 uM ionomycin for 5 min were fixed and imaged by confocal microscopy. (C) Indicated proteins were detected by SDS-PAGE and Western blotting of lysates with Munc13-4 antibody from MDA-MB-231 cells stably expressing control shRNA (Ctrl) or shRNA targeting Munc13-4 (KD), or Munc13-4 KD cells rescued with shRNA-resistant wild-type Munc13-4, Munc13-4 C2A*, or Munc13-4 C2B*. (D) Culture media supernatants (as in Fig. 1 B) from MDA-MB-231 cells as in C either untreated or stimulated with 1.25 uM ionomycin for 30 min were filtered onto membrane and analyzed for CD63 or GM130. (E) Quantification of CD63+ exosome release (from Fig 1 D) shown as mean values +/- SE for n = 3. *, P < 0.05 for comparison between ionomycin-treated and basal. Bars, 5 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29930202), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for UNC13D/Munc 13-4 Antibody

Application
Recommended Usage

Immunohistochemistry

2.5 ug/mL

Immunohistochemistry-Paraffin

2.5 ug/mL

Peptide ELISA

Detection limit 1:128000

Western Blot

1 - 3 ug/mL
Application Notes
WB: Approx. 105 kDa band observed in human T-lymphocyte and HeLa lysates (calculated MW of 123 kDa band according to NP_954712.1). IHC-P: Human thymus shows cytoplasm staining in select areas. Mouse Lung shows staining of the resperatory epithelium in wildtype but not in the knock-out, provided by Micheal Tuvim, University of Texas MD Anderson Cancer Center, Houston, USA. Use in ICC/IF reported in scientific literature (PMID 28100639).

Reviewed Applications

Read 1 review rated 5 using NB100-41385 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

Tris saline (20 mM Tris pH 7.3, 150 mM NaCl), 0.5% BSA

Preservative

0.02% Sodium Azide

Concentration

0.5 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C. Avoid freeze-thaw cycles.

Background: UNC13D

This gene encodes a protein that is a member of the UNC13 family, containing similar domain structure as other family members but lacking an N-terminal phorbol ester-binding C1 domain present in other Munc13 proteins. The protein appears to play a role in vesicle maturation during exocytosis and is involved in regulation of cytolytic granules secretion. Mutations in this gene are associated with familial hemophagocytic lymphohistiocytosis type 3, a genetically heterogeneous, rare autosomal recessive disorder.

Long Name

UNC-13 Homolog D

Alternate Names

FHL3, HLH3, HPLH3, Munc13-4

Entrez Gene IDs

201294 (Human); 70450 (Mouse); 192177 (Rat)

Gene Symbol

UNC13D

UniProt

NP_954712.1 (Human)

Additional UNC13D Products

Product Documents for UNC13D/Munc 13-4 Antibody

Certificate of Analysis

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

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Product Specific Notices for UNC13D/Munc 13-4 Antibody

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Related Research Areas

Lysosomal Enzymes

Citations for UNC13D/Munc 13-4 Antibody

Customer Reviews for UNC13D/Munc 13-4 Antibody (1)

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Customer Images

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  • UNC13D/Munc 13-4 Antibody
    Name: Scott Messenger
    Application: Western Blot
    Sample Tested: RBL-2H3 rat basophilic leukemia cell line
    Species: Rat
    Verified Customer | Posted 04/14/2017
    RBL-2H3 from Wt or shRNA Munc13-4 lentiviral stable expression whole cell lysate. 50ug protein
    UNC13D/Munc 13-4 Antibody NB100-41385

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Protocols

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FAQs for UNC13D/Munc 13-4 Antibody

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  • Q: I'm looking Munc13-4 N terminal antibody with related epitope sequence.

    A:

    Unfortunately we do not epitope map our antibodies, but we do have a whole list of antibodies against your target depending on species reactivity and application you want to use them for. Please see this link to our MUNC 13-4 antibodies. All of the immunogen information that is not proprietary, or known should be presented on the datasheet. Often times a range will be provided where the peptide falls within, or if it was raised against a larger recombinant protein as is the case for the one you inquired about we do not know exactly where the binding occurs.

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