Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Western Blot, Immunocytochemistry

Cited:

Immunohistochemistry, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG
View all ASK1 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human ASK1
Lys1011-Asp1196
Accession # Q99683

Specificity

Detects human ASK1 in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Human ASK1 Antibody

Detection of Human ASK1 antibody by Western Blot.

Detection of Human ASK1 by Western Blot.

Western blot shows lysates of Raji human Burkitt's lymphoma cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human ASK1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3575) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for ASK1 at approximately 154 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

Detection of Human ASK1 by Western Blot

Detection of Human ASK1 by Western Blot

M. tb virulence induces ASK1 expression. Human MDMs were obtained after seven days in culture. 2 × 106 MDMs were infected at MOI 1 and MOI 10 with an avirulent (H37Ra) and MOI 1 and MOI 5 with a virulent (H37Rv) strain of M. tb; 2 × 106 MDMs were not infected as a control (Uninf). At 24 h postinfection, cells were recovered and prepared for Western Blot. Representative Western blot of ASK1, JNK1, JNK2, p-p38, and GAPDH (a). Band densities of ASK1 (b), JNK1 (c), JNK2 (d), and p-p38 (e) were normalized against GAPDH and quantified by densitometry analysis with the ImageJ software. Results are shown in relative units (RU) of concentration. The bar graphs show the mean ± SD from two independent experiments (n = 2 donors and two technical replicates). Statistical analysis was performed using Kruskal–Wallis analysis, followed by Dunn’s post hoc test. * p < 0.05. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35631013), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human ASK1 by Western Blot

Detection of Human ASK1 by Western Blot

M. tb virulence induces ASK1 expression. Human MDMs were obtained after seven days in culture. 2 × 106 MDMs were infected at MOI 1 and MOI 10 with an avirulent (H37Ra) and MOI 1 and MOI 5 with a virulent (H37Rv) strain of M. tb; 2 × 106 MDMs were not infected as a control (Uninf). At 24 h postinfection, cells were recovered and prepared for Western Blot. Representative Western blot of ASK1, JNK1, JNK2, p-p38, and GAPDH (a). Band densities of ASK1 (b), JNK1 (c), JNK2 (d), and p-p38 (e) were normalized against GAPDH and quantified by densitometry analysis with the ImageJ software. Results are shown in relative units (RU) of concentration. The bar graphs show the mean ± SD from two independent experiments (n = 2 donors and two technical replicates). Statistical analysis was performed using Kruskal–Wallis analysis, followed by Dunn’s post hoc test. * p < 0.05. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35631013), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human ASK1 Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed human peripheral blood mononuclear cells

Western Blot

1 µg/mL
Sample: Raji human Burkitt's lymphoma cell line

Reviewed Applications

Read 1 review rated 4 using AF3575 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 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: ASK1

Apoptosis signal-regulating kinase 1 (ASK1) is a MAP kinase kinase kinase also known as MEKK5 and MAP3K5. ASK1 is induced by inflammatory cytokines, UV light, and other stress-related stimuli to phosphorylate MAP kinase kinases that in turn activate the p38 and JNK families of MAP kinases. Mice lacking ASK1 are resistant to stress-induced p38 and JNK activation and cell death.

Long Name

Apoptosis Signal-regulated Kinase 1

Alternate Names

MAP3K5, MEKK5

Entrez Gene IDs

4217 (Human); 26408 (Mouse)

Gene Symbol

MAP3K5

UniProt

Q99683

Additional ASK1 Products

Product Documents for Human ASK1 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human ASK1 Antibody

For research use only

Citations for Human ASK1 Antibody

Customer Reviews for Human ASK1 Antibody (1)

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  • Human ASK1 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: MDA-MB-231 human breast cancer cell line
    Species: Human
    Verified Customer | Posted 01/25/2018
    Human ASK1 Antibody AF3575

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Protocols

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