- PA1-46489 - Invitrogen Antibodies S100A9 antibody

Submitted references Rapamycin-filgrastim combination therapy ameliorates portal hypertension-induced splenomegaly: Role of β actin and S100A9 proteins modulation.

Abdelrahman SA, Abdelfatah MM, Keshta AT
Iranian journal of basic medical sciences 2022 Jun;25(6):732-744

S100A8 and S100A9, both transcriptionally regulated by PU.1, promote epithelial-mesenchymal transformation (EMT) and invasive growth of dermal keratinocytes during scar formation post burn.

Xu Z, Cheng C, Kong R, Liu Y, Wang S, Ma Y, Xing X
Aging 2021 Jun 7;13(11):15523-15537

Quantitative proteomic characterization of microvesicles/exosomes from the cerebrospinal fluid of patients with acute bilirubin encephalopathy.

Tan N, Hu S, Hu Z, Wu Z, Wang B
Molecular medicine reports 2020 Aug;22(2):1257-1268

Distinct molecular signatures of mild extrinsic and intrinsic atopic dermatitis.

Martel BC, Litman T, Hald A, Norsgaard H, Lovato P, Dyring-Andersen B, Skov L, Thestrup-Pedersen K, Skov S, Skak K, Poulsen LK
Experimental dermatology 2016 Jun;25(6):453-9

Analysis of protein expression in periodontal pocket tissue: a preliminary study.

Monari E, Cuoghi A, Bellei E, Bergamini S, Lucchi A, Tomasi A, Cortellini P, Zaffe D, Bertoldi C
Proteome science 2015;13:33

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of S100A9 Antibody in DMSO treated HL60 whole cell lysates.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of S100A9 in 5 µg /lane cells transfected with the pCMV6-ENTRY control or pCMV6-ENTRY S100A9 cDNA. Samples were incubated in S100A9 polyclonal antibody (Product # PA1-46489). Cells were transfected with the pCMV6-ENTRY control or pCMV6-ENTRY S100A9 cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates were separated by SDS-PAGE and immunoblotted.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of S100A9 in DMSO treated HL60 whole cell lysates. Sample was incubated in S100A9 polyclonal antibody (Product # PA1-46489).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of S100A9 in 0.5 mg/mL Human PBMCs lysate. Samples were incubated in S100A9 polyclonal antibody (Product # PA1-46489). This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-S100A9 Polyclonal Antibody (Product # PA1-46489) and a 13kDa band corresponding to S100A9 was observed in SK-BR-3, THP-1, Mouse Spleen and Rat Spleen but was absent in Raji, SH-SY5Y, Mouse Brain and Rat Brain which are reported to be negative. Expression of Calprotectin S100A9 was downregulated upon treatment of THP-1 with PMA/TPA [10.1073/pnas.0709958105]. Membrane enriched cell extracts (30 µg lysate) of SK-BR-3 (Lane 1), THP-1 (Lane 2), THP-1 treated with PMA/TPA (50ng/ml for 24 hours) (Lane 3), Raji (Lane 4), SH-SY5Y (Lane 5), Mouse Spleen (Lane 6), Rat Spleen (Lane 7), Mouse Brain (Lane 8) and Rat Brain (Lane 9) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of S100A9 was performed using log phase THP-1 and Raji cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with S100A9 Polyclonal Antibody (Product # PA1-46489) at 1:200 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing localization to plasma membrane, nucleus and cytoplasm. Panel e shows Raji cells with no expression of S100A9. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Histological analysis of proteins involved in epidermal barrier function. Immunohistochemical staining (brown) of filaggrin, loricrin, Ki-67 and S100A9 in skin biopsies from patients with extrinsic atopic dermatitis ( AD ex), intrinsic AD ( AD in) and healthy controls ( NN ). (a) Filaggrin staining of skin samples from NN and patients with a detected mutation in FLG at the indicated loci: 2282del4, R2447x or R501x. (b) Loricrin staining of skin samples from NN and patients with a mutation in FLG . (c) Ki-67 staining of skin samples from NN and patients with extrinsic and intrinsic AD , respectively. (d) S100A9 staining of skin samples from NN and patients with extrinsic and intrinsic AD , respectively. Numbers correspond to the ID of the specific subjects as in Table S1. Scale bars = 250 mu m.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 3 Detailed protein spots, differentially expressed in C and T specimens

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 4 Representative images of Western blot analyses in C and T specimens for LEG7, 14-3-3, HSPB1, S100A9 and beta-actin. The beta-actin was utilized as reference for protein samples loading and integrity

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 6. Verification of differentially expressed proteins using western blotting. (A) Representative western blots demonstrate the expression levels of LTF, DEFA1, S100A7 and S100A9 in the MV/E isolated from the CSF. Histogram data shows the densitometric semi-quantification (mean +- SD) of (B) LTF, (C) DEFA1, (D) S100A7 and (E) S100A9 expression levels. Protein densitometric semi-quantification was normalized to the density of CD9 protein. CD9 is an exosome surface marker. *P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 10 S100A9 immunostained section in the spleen showing A: Control group shows positive cytoplasmic immunoreaction for S100 antibody in some cells of the red pulp (arrow). B: TAA group showing increased cytoplasmic immunoreaction in cells of red and white pulp (arrow). C: Rapamycin-treated group showing increased cytoplasmic immunoreaction in cells of red pulp (arrow). D: Filgrastim-treated group showing positive cytoplasmic immunoreaction in some cells of the red pulp (arrow). E: Combined rapamycin and filgrastim-treated group showing positive reaction in few cells of red pulp (arrow) (Immunoperoxidase technique for S100 x 40, scale bar 30 um) TTA: thioacetamide

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Different expression of proteins involved in epidermal barrier function. (a, b) Filaggrin and loricrin coverage, respectively. Coverage was determined manually by subtracting the length of the unstained part of the epidermis from the entire length of the epidermis. (c) Correlation of filaggrin and loricrin coverage. (d) Intensity of S100A9 staining multiplied by area and normalized to the length of the skin biopsy. (e) Area of Ki-67 staining normalized to the length of the basal layer. n ( AD ex) = 5; n ( AD in) = 8; n (Pso) = 8; n ( NN ) = 8. Horizontal bars: mean with SD . * P < 0.05; *** P < 0.001. Comparison of groups: 1-way ANOVA with Tukey's multiple comparison test.

PA1-46489

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