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Collagen III Recombinant Rabbit Monoclonal Antibody [JE60-79] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA751826

Collagen III Recombinant Rabbit Monoclonal Antibody [JE60-79] - BSA and Azide free

Application

  • WB

  • IF-Cell

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

Collagen III Recombinant Rabbit Monoclonal Antibody [JE60-79] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human Collagen III 101-400/1,466.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, FC

Molecular Weight

Predicted band size: 139 kDa

Positive Control

HeLa cell lysate, A431 cell lysate, SiHa cell lysate, HaCaT cell lysate, NIH/3T3 cell lysate, RAW264.7 cell lysate, C6 cell lysate, PC-12 cell lysate, Mouse heart tissue lysate, Mouse skin tissue lysate, Mouse skeletal muscle tissue lysate, Rat skeletal muscle tissue lysate, Rat skin tissue lysate, SiHa, PC-12, HeLa.

Conjugation

unconjugated

Clone Number

JE60-79

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:2,000

  • IF-Cell

  • 1:100-1:500

  • FC

  • 1:500-1:1,000

Target

Function

Collagen type III occurs in most soft connective tissues along with type I collagen. Involved in regulation of cortical development. Is the major ligand of ADGRG1 in the developing brain and binding to ADGRG1 inhibits neuronal migration and activates the RhoA pathway by coupling ADGRG1 to GNA13 and possibly GNA12. This gene encodes the pro-alpha1 chains of type III collagen, a fibrillar collagen that is found in extensible connective tissues such as skin, lung, uterus, intestine and the vascular system, frequently in association with type I collagen. Mutations in this gene are associated with Ehlers-Danlos syndrome types IV, and with aortic and arterial aneurysms. Two transcripts, resulting from the use of alternate polyadenylation signals, have been identified for this gene.

Background References

1. Wang R. et. al. Long noncoding RNA DNM3OS promotes prostate stromal cells transformation via the miR-29a/29b/COL3A1 and miR-361/TGFbeta1 axes. Aging (Albany NY). 2019 Nov

2. Kuivaniemi H. et. al. Type III collagen (COL3A1): Gene and protein structure, tissue distribution, and associated diseases. Gene. 2019 Jul

Subcellular Location

Extracellular matrix.

UNIPROT

SwissProt: P02461 Human

SwissProt: P08121 Mouse

SwissProt: P13941 Rat

Synonyms

Alpha 1 type III collagen antibody

Alpha1 (III) collagen antibody

CO3A1_HUMAN antibody

COL 3A1 antibody

COL3A1 antibody

Collagen alpha 1(III) chain antibody

Collagen alpha-1(III) chain antibody

Collagen III alpha 1 chain precursor antibody

Collagen III alpha 1 polypeptide antibody

Collagen type III alpha 1 (Ehlers Danlos syndrome type IV autosomal dominant) antibody

Expand

Images

  • Western blot analysis of Collagen III on different lysates with Rabbit anti-Collagen III antibody (<a href="/products/HA751826" style="font-weight: bold;text-decoration: underline;">HA751826</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: A431 cell lysate (20 µg/Lane)<br />Lane 3: SiHa cell lysate (20 µg/Lane)<br />Lane 4: HaCaT cell lysate (20 µg/Lane)<br />Lane 5: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 6: RAW264.7 cell lysate (20 µg/Lane)<br />Lane 7: C6 cell lysate (20 µg/Lane)<br />Lane 8: PC-12 cell lysate (20 µg/Lane)<br />Lane 9: Mouse heart tissue lysate (20 µg/Lane)<br />Lane 10: Mouse skin tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 139 kDa<br />Observed band size: 150 kDa<br /><br />Exposure time: 2 seconds; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA751826" style="font-weight: bold;text-decoration: underline;">HA751826</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Collagen III on different lysates with Rabbit anti-Collagen III antibody (HA751826) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: A431 cell lysate (20 µg/Lane)
    Lane 3: SiHa cell lysate (20 µg/Lane)
    Lane 4: HaCaT cell lysate (20 µg/Lane)
    Lane 5: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 6: RAW264.7 cell lysate (20 µg/Lane)
    Lane 7: C6 cell lysate (20 µg/Lane)
    Lane 8: PC-12 cell lysate (20 µg/Lane)
    Lane 9: Mouse heart tissue lysate (20 µg/Lane)
    Lane 10: Mouse skin tissue lysate (20 µg/Lane)

    Predicted band size: 139 kDa
    Observed band size: 150 kDa

    Exposure time: 2 seconds; ECL: K1801;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751826) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of Collagen III on different lysates with Rabbit anti-Collagen III antibody (<a href="/products/HA751826" style="font-weight: bold;text-decoration: underline;">HA751826</a>) at 1/1,000 dilution.<br /><br />Lane 1: Mouse heart tissue lysate (20 µg/Lane)<br />Lane 2: Mouse skeletal muscle tissue lysate (20 µg/Lane)<br />Lane 3: Rat skeletal muscle tissue lysate (20 µg/Lane)<br />Lane 4: Rat skin tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 139 kDa<br />Observed band size: 150 kDa<br /><br />Exposure time: 2 minutes; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA751826" style="font-weight: bold;text-decoration: underline;">HA751826</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Collagen III on different lysates with Rabbit anti-Collagen III antibody (HA751826) at 1/1,000 dilution.

    Lane 1: Mouse heart tissue lysate (20 µg/Lane)
    Lane 2: Mouse skeletal muscle tissue lysate (20 µg/Lane)
    Lane 3: Rat skeletal muscle tissue lysate (20 µg/Lane)
    Lane 4: Rat skin tissue lysate (20 µg/Lane)

    Predicted band size: 139 kDa
    Observed band size: 150 kDa

    Exposure time: 2 minutes; ECL: K1801;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751826) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of SiHa cells labeling Collagen III with Rabbit anti-Collagen III antibody (<a href="/products/HA751826" style="font-weight: bold;text-decoration: underline;">HA751826</a>) at 1/500 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Collagen III antibody (<a href="/products/HA751826" style="font-weight: bold;text-decoration: underline;">HA751826</a>) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of SiHa cells labeling Collagen III with Rabbit anti-Collagen III antibody (HA751826) at 1/500 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Collagen III antibody (HA751826) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunocytochemistry analysis of PC-12 cells labeling Collagen III with Rabbit anti-Collagen III antibody (<a href="/products/HA751826" style="font-weight: bold;text-decoration: underline;">HA751826</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Collagen III antibody (<a href="/products/HA751826" style="font-weight: bold;text-decoration: underline;">HA751826</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of PC-12 cells labeling Collagen III with Rabbit anti-Collagen III antibody (HA751826) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Collagen III antibody (HA751826) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Flow cytometric analysis of HeLa cells labeling Collagen III.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA751826" style="font-weight: bold;text-decoration: underline;">HA751826</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HeLa cells labeling Collagen III.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA751826, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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