14-9771-80

antibody from Invitrogen Antibodies

Targeting: COL10A1

Provider product page for 14-9771-80

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Immunohistochemistry

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Antibody data

Antibody Data
Antigen structure
References [17]
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Validations
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Product number: 14-9771-80 - Provider product page
Provider: Invitrogen Antibodies
Product name: Collagen X Monoclonal Antibody (X53), eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: The monoclonal antibody X53 recognizes human, mouse, porcine, guinea pig, and rat Collagen Type X. X53 does not cross-react with collagen types I, II, III, IV, or VI. Collagen Type X is a homotrimeric short chain collagen. Collagen Type X has limited expression in normal adult tissues, primarily being expressed during bone growth and development. Expression is localized to the peri- and extracellular matrix of hypertrophic chondrocytes during endochondral ossification and also plays a role in cartilage mineralization. Collagen Type X is elevated in many tumor types with expression being localized in the vasculature. Mutations to the gene COL10A1, which encodes human Collagen Type X, is associated with Schmid type metaphyseal chondrodysplasia. Applications Reported: This X53 antibody has been reported for use in microscopy, immunohistochemical staining, ELISA, and immunocytochemistry. Applications Tested: This X53 antibody has been tested by immunohistochemistry on formalin-fixed paraffin embedded human tissue using low pH antigen retrieval and can be used at less than or equal to 10 µg/mL. 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, Mouse, Rat, Canine, Guinea Pig, Porcine
Host: Mouse
Isotype: IgG
Antibody clone number: X53
Vial size: 25 µg
Concentration: 0.5 mg/mL
Storage: 4° C

COL10A1 protein structure - 14-9771-80 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 7 In vivo osteochondral repair in a mouse model: evaluation of chondrogenic repair ( a ) Histology of the tissue stained with toluidine blue (bar = 500 um). ( b ) Histology of the tissue stained with Saf-O (left columns; bar = 500 um) and the corresponding high-magnification images (right columns; bar = 100 um). ( c ) Immunohistochemical analysis of repair cartilage stained for COL2 (bar = 500 um). ( d ) Immunohistochemical analysis of repair tissue stained for COL10 (bar = 500 um). bFGF, basic fibroblast growth factor; Saf-O, Safranin-O; COL2, collagen type II; COL10, collagen type X.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 Hypertrophic cartilage area and number of TRAP-positive callus cells . ( A , B ) Number/area of hypertrophic chondrocytes, visualized by collagen X staining, in Tac1-/-, alpha-CGRP-/- ( A ), SYX ( B ), and WT mice ( A , B ) at the time points 5, 9, 13, 16, and 21 days after fracture. Values were calculated as a percentage of total callus area and are presented as mean +- SD. * in black indicates differences between neurotransmitter deficient mice and WT animals. ( C , D ) Number of TRAP-positive cells in the callus tissue of Tac1-/-, alpha-CGRP-/- ( C ), SYX ( D ), and WT mice ( C , D ) at the time points 5, 9, 13, 16, and 21 days after fracture. The number of cells was calculated in relation to the total area of callus tissue as number/mm 2 . ( E ) Representative image of collagen X stained hypertrophic callus area of an alpha-CGRP-/- mouse 13 days after fracture. ( F ) Representative image of TRAP-positive callus cells 16 days after fracture in the callus tissue of a WT mouse. Overview images scanned with 20x magnification (TissueFAXSi plus). Red boxes demonstrate the enlarged view. Red arrow points to stained cells. Col X = collagen X; B = bone, C = Callus tissue, F = Fracture site, M = Muscle. * = p

Supportive validation

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Experimental details: FIGURE 1 Chondrogenic differentiation of bone marrow derived mesenchymal stem cells (BMSC). (A) Histological and Immunohistological staining of day 21 chondrogenically differentiated BMSC-pellets of the donors for Collagen type II, Thionine and Collagen type X. (B) qPCR Gene-expression analysis of chondrocyte and hypertrophy marker genes in day 21 chondrogenically differentiated BMSCs. Raw average Ct value for COL2A1 = 20.54; COL10A1 = 20.99; ALPL = 28.19.

Supportive validation

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Experimental details: Figure 5 Effects of KAG-308 on chondrocyte hypertrophy in vitro . ( a ) mRNA levels of Col10a1 , Runx2 , Mef2c , and Col2a1 in pellet culture of mouse articular chondrocytes treated with indicated concentration of KAG-308 for 2 wks. Symbols represent individual pellets; long and short bars show the mean and SD, respectively. * P < 0.05, ** P < 0.005, *** P < 0.0005 vs vehicle by ANOVA followed by Dunnett's post hoc test. ( b ) Safranin O staining and Col10a1 immunostaining of cultured pellets of mouse articular chondrocytes treated with KAG-308 for 2 wks. Inset boxes in safranin-O staining indicate enlarged images. Scale bar, 200 um. ( c ) Immunoblotting of Hdac4 in nuclear and cytosolic fractions from mouse articular chondrocytes in a time course with 10 nM KAG-308 treatment. Right graphs indicate quantitative densitometry analysis of the immunoblots (n = 5). Hdac4 values were normalized to Pcna (nuclear) or Actin (cytosol). Long and short bars show the mean and SD, respectively. * P < 0.05, ** P < 0.005 vs 0 hour by ANOVA followed by Dunnett's post hoc test. ( d ) Immunocytochemistry of Hdac4 in mouse articular chondrocytes after 4-d-culture in BMP-2-containing medium with vehicle or 10 nM KAG-308. Right graph shows the percent of cells with nuclear-localized Hdac4. Symbols represent independent slides; long and short bars show the mean and SD, respectively. * P < 0.0005 by Welch's t test. Scale bar, 20 um.

Supportive validation

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Experimental details: Figure 5 Accelerated chondrogenesis in SHIP-1 KO PalphaS MSCs. PalphaS MSC pellets differentiated in vitro for 21 days using the Mouse StemXVivo Chondrogenic Base Media and Supplement were fixed, paraffin embedded, sectioned and IHC-stained. (a and b) Sections from WT (a) and SHIP-1 KO (b) PalphaS MSC pellets were stained with an antibody against the early chondrogenic lineage marker Col II. Bound antibody was visualized in bright red with a NorthernLights 557-conjugated anti-sheep secondary antibody. (c and d) Sections from WT (c) and SHIP-1 KO (d) PalphaS MSC pellets were stained with a bright red eFluor570-conjugated antibody against the hypertrophic chondrocyte lineage marker Col X. (e and f) Sequential sections from the same WT cell pellet as in (c) and from the same SHIP-1 KO cell pellet as in (d) were subjected to H&E staining. Magnification: x200 (a-d); x400 (e and f). Scale bar = 50 mum. (g) Left panel, the difference in cell sizes between WT and SHIP-1 KO PalphaS MSCs after chondrogenic induction was quantified by using the NIH ImageJ program and indicated as ImageJ Area Units. Middle and right panels, gene expression levels of Col2a1 and Col10a1 were evaluated by quantitative real-time PCR. GAPDH expression was assessed as an internal reference for quantification. Gene expression levels were expressed as relative fold increase over the internal control. Student's t test was performed and p < .05 is considered significant. Col II, collagen type II; Col X, collagen t

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 8 Immunofluorescence images illustrating deep and surface sections. Staining of collagen I (red), collagen II (green), aggrecan (green) and collagen X (green) was performed on constructs from all groups on day 28 (as indicated; DAPI nuclei staining in light blue); ( A , E , I ): GelMA-ALG; ( B , F , J ): GelMA/HAMA-ALG; ( C , G , K ): GelMA-ALG/HAP and ( D , H , L ): GelMA/HAMA-ALG/HAP. Images were taken from the deep hydrogel zone near to the ALG or ALG/HAP scaffold, with surface images inset; scale bar 50 um.