14-7118-85

antibody from Invitrogen Antibodies

Targeting: IL1A IL-1A, IL1, IL1-ALPHA, IL1F1

ELISA (Recommended by provider)

Flow cytometry (Recommended by provider)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

14-7118-85

Provider

Invitrogen Antibodies

Product name

IL-1 alpha Monoclonal Antibody (364/3B3-14), eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Description: The 364/3B3-14 antibody reacts with human interleukin-1alpha. Applications Reported: The 364/3B3-14 antibody has been reported for intracellular staining for flow cytometric analysis. Applications Tested: This 364/3B3-14 antibody has been tested intracellular staining and flow cytometric analysis of stimulated human blood cells. This can be used at less than or equal to 1 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC. Filtration: 0.2 µm post-manufacturing filtered.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

364/3B3-14

Vial size

500 µg

Concentration

0.5 mg/mL

Storage

4° C

References

Submitted references

DOT1L modulates the senescence-associated secretory phenotype through epigenetic regulation of IL1A.

Leon KE, Buj R, Lesko E, Dahl ES, Chen CW, Tangudu NK, Imamura-Kawasawa Y, Kossenkov AV, Hobbs RP, Aird KM
The Journal of cell biology 2021 Aug 2;220(8)

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Ultraviolet B radiation-induced cell death: critical role of ultraviolet dose in inflammation and lupus autoantigen redistribution.

Caricchio R, McPhie L, Cohen PL
Journal of immunology (Baltimore, Md. : 1950) 2003 Dec 1;171(11):5778-86

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Dendritic cell activation and cytokine production induced by group B Neisseria meningitidis: interleukin-12 production depends on lipopolysaccharide expression in intact bacteria.

Dixon GL, Newton PJ, Chain BM, Katz D, Andersen SR, Wong S, van der Ley P, Klein N, Callard RE
Infection and immunity 2001 Jul;69(7):4351-7

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Sensitive and specific immunoradiometric assays for human interleukin-1 alpha.

Thorpe R, Wadhwa M, Gearing A, Mahon B, Poole S
Lymphokine research 1988 Summer;7(2):119-27

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 2. DOT1L is necessary for H3K79me2/3 at the IL1A locus and SASP expression but dispensable for other senescence phenotypes. IMR90 cells were infected with retrovirus-expressing HRAS G12V (RAS) or empty vector control with or without shRNA to human DOT1L (shDOT1L) or an shGFP control. Details on time points are in Fig. S1 A . (A) Immunoblot analysis of total cell lysates (TCL) and chromatin fractions of the indicated proteins. beta-Actin was used as a loading control for TCL. Histone H3 was used as a loading control for chromatin fractions. One of three independent experimental replicates is shown. (B) H3K79me2 and DOT1L binding to the IL1A promoter region and H3K79me2 at the promoters of IL6 , CXCL8 , and IL1B was determined by ChIP-qPCR and normalized to total histone H3 binding at the same site. One of three independent experimental replicates is shown. Data represent mean +- SD ( n = 3). *, P < 0.05 by one-way ANOVA with Tukey's multiple comparisons. (C) H3K79me3 and DOT1L binding to the IL1A gene body and H3K79me3 at the gene bodies of IL6 , CXCL8 , and IL1B was determined by ChIP-qPCR and normalized to total histone H3 binding at the same site. One of three independent experimental replicates is shown. Data represent mean +- SD ( n = 3). *, P < 0.05 by one-way ANOVA with Tukey's multiple comparisons. (D) IL1A mRNA expression was determined by RT-qPCR. One of five independent experimental replicates is shown. Data represent mean +- SD ( n = 3). *, P < 0.01 by one-w

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 4. DOT1L overexpression (OE) increases H3K79me2/3 at the IL1A locus and is sufficient for SASP gene expression but does not affect other senescence phenotypes. IMR90 cells were infected with retrovirus-expressing human DOT1L or empty vector control. In some experiments, as a positive control, IMR90 cells were infected with retrovirus-expressing HRAS G12V (RAS) or empty vector control. (A) DOT1L mRNA expression. One of three independent experimental replicates is shown. Data represent mean +- SD ( n = 3). *, P < 0.001 by Student's t test. (B) DOT1L immunoblot analysis. Vinculin was used as a loading control. One of five independent experimental replicates is shown. (C) H3K79me2 and H3K79me3 immunoblot analysis on chromatin fractions. Total histone H3 was used as loading control. One of three independent experimental replicates is shown. (D) H3K79me2 and DOT1L binding to the IL1A promoter region and H3K79me2 at the promoters of IL6 , CXCL8 , and IL1B was determined by ChIP-qPCR and normalized to total histone H3 binding at the same site. One of three independent experimental replicates is shown. Data represent mean +- SD ( n = 3). *, P < 0.05 by Student's t test. (E) H3K79me3 and DOT1L binding to the IL1A gene body and H3K79me3 at the gene bodies of IL6 , CXCL8 , and IL1B was determined by ChIP-qPCR and normalized to total histone H3 binding at the same site. One of three independent experimental replicates is shown. Data represent mean +- SD ( n = 3). *, P < 0.05 by Stu