Anti-Thrombin antibody [5G9]

• IHC-P

AbReview41773****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombin antibody [5G9] (AB17199)

ab17199 staining Thrombin in human liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde, permeabilized with 0.2% Triton X-100 in PBS and blocked with 5% milk for 30 minutes at room temperature; antigen retrieval was by heat mediation in Tris pH 9.0. Samples were incubated with primary antibody (1/100 in PBS) for 16 hours at 4°C. An undiluted Biotin-conjugated horse anti-mouse IgG polyclonal was used as the secondary antibody.

This image is courtesy of an Abreview submitted by Steffen Rickelt

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombin antibody [5G9] (AB17199)

IHC image of Thrombin staining in human liver formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab17199, 10μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• WB

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Western blot - Anti-Thrombin antibody [5G9] (AB17199)

Thrombin contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.

All lanes:

Western blot - Anti-Thrombin antibody [5G9] (ab17199) at 1 µg/mL

Lane 1:

Human liver tissue lysate - total protein (<a href='/en-us/products/unavailable/human-liver-tissue-lysate-total-protein-ab29889'>ab29889</a>) at 10 µg

Lane 2:

Human Plasma Total Protein Lysate at 10 µg

Lane 3:

Western blot - Hep G2 nuclear extract lysate (<a href='/en-us/products/tissue-lysates/hep-g2-nuclear-extract-lysate-ab14660'>ab14660</a>) at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-hrp-preadsorbed-ab97040'>ab97040</a>) at 1/5000 dilution

Predicted band size: 70 kDa

Observed band size: 40 kDa,75 kDa

true

Exposure time: 12min

• WB

CiteAb

Western blot - Anti-Thrombin antibody [5G9] (AB17199)

Thrombin western blot using anti-Thrombin antibody [5G9] ab17199. Publication image and figure legend from Tang, X., Zhang, Z., et al., 2020, Cell Res, PubMed 31811276.

ab17199 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab17199 please see the product overview.

Effects of both apo- and holo-transferrin on thrombin, FXIIa and antithrombin. a Potentiating effects of both apo- and holo-transferrin on thrombin. b, c Representative RP-HPLC analysis (b) and quantification (c) of fibrinopeptide A (FbpA) and fibrinopeptide B (FbpB) released from 5 mg of fibrinogen hydrolyzed by 0.1 NIH unit thrombin mixed with 0, 0.2, 1, or 5 μM apo-transferrin, respectively. d Potentiating effects of both apo- and holo-transferrin on FXIIa. e, f Representative western blot (e) and quantification analysis of kallikrein heavy chain (HC ~52 kDa) (f) released from 10 μg of prekallikrein (PK) hydrolyzed by 0.01 NIH unit FXIIa mixed with 0, 0.2, 1, or 5 μM apo-transferrin (lane 2–5), respectively. Blots of PK, FXIIa heavy chain (FXIIa HC), transferrin, and kallikrein light chain (LC ~36 and 33 kDa) are also shown. g–j Apo- and holo-transferrin block antithrombin (AT)'s inactivation effect on thrombin (g, h) and FXa (i, j). TAT : thrombin–AT complex. HSA : human serum albumin. Data represent mean ± SD of five independent experiments, *p < 0.05, **p < 0.01 by one-way ANOVA with Dunnett's post hoc test. N.S. : no significance; Tf : transferrin

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