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c-Kit Recombinant Rabbit Monoclonal Antibody [ST04-99]

Usd: 385

Specification

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Catalog# ET1609-60

c-Kit Recombinant Rabbit Monoclonal Antibody [ST04-99]

Application

  • WB

  • IHC-P

Reactivity

  • Human

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

c-Kit Recombinant Rabbit Monoclonal Antibody [ST04-99]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within C-terminal human KIT.

Species Reactivity

Human

Validated Applications

WB, IHC-P

Molecular Weight

Predicted band size: 110 kDa

Positive Control

Human brain tissue lysates, Saos-2 cell lysate, human breast tissue, human gastrointestinal stromal tumor tissue.

Conjugation

unconjugated

Clone Number

ST04-99

RRID

AB_3069869

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IHC-P

  • 1:200

Target

Function

The c-Kit proto-oncogene is a member of the receptor tyrosine kinase family and, more specifically, is closely related to the platelet derived growth factor receptor (PDGFR). c-Kit, the normal cellular homolog of the HZ4-feline sarcoma virus transforming gene (v-Kit), encodes a transmembrane receptor. c-Kit regulates a variety of biological responses including chemotaxis, cell proliferation, apoptosis and adhesion. c-Kit is also identical with the product of the W locus in mice and, as such, is integral to the development of mast cells and hematopoiesis. The ligand for the c-Kit receptor (KL) has been identified and is encoded at the murine steel (SI) locus. Kit is the human homolog of the proto-oncogene c-Kit. Mutations in Kit are integral for tumor growth and progression in various cancers.

Background References

1. Chi C et al. In vitro morphology, viability and cytokine secretion of uterine telocyte-activated mouse peritoneal macrophages. J Cell Mol Med 19:2741-50 (2015).

2. Drummond CA et al. Reduction of Na/K-ATPase affects cardiac remodeling and increases c-kit cell abundance in partial nephrectomized mice. Am J Physiol Heart Circ Physiol 306:H1631-43 (2014).

Sequence Similarity

Belongs to the protein kinase superfamily. Tyr protein kinase family. CSF-1/PDGF receptor subfamily.

Tissue Specificity

Isoform 1 and isoform 2 are detected in spermatogonia and Leydig cells. Isoform 3 is detected in round spermatids, elongating spermatids and spermatozoa (at protein level). Widely expressed. Detected in the hematopoietic system, the gastrointestinal system, in melanocytes and in germ cells.

Post-translational Modification

Ubiquitinated by SOCS6. KIT is rapidly ubiquitinated after autophosphorylation induced by KITLG/SCF binding, leading to internalization and degradation.; Autophosphorylated on tyrosine residues. KITLG/SCF binding enhances autophosphorylation. Isoform 1 shows low levels of tyrosine phosphorylation in the absence of added KITLG/SCF (in vitro). Kinase activity is down-regulated by phosphorylation on serine residues by protein kinase C family members. Phosphorylation at Tyr-568 is required for interaction with PTPN11/SHP-2, CRK (isoform Crk-II) and members of the SRC tyrosine-protein kinase family. Phosphorylation at Tyr-570 is required for interaction with PTPN6/SHP-1. Phosphorylation at Tyr-703, Tyr-823 and Tyr-936 is important for interaction with GRB2. Phosphorylation at Tyr-721 is important for interaction with PIK3R1. Phosphorylation at Tyr-823 and Tyr-936 is important for interaction with GRB7.

Subcellular Location

Cell membrane, Cytoplasm.

UNIPROT

SwissProt: P10721 Human

SwissProt: P05532 Mouse

Synonyms

C Kit antibody

c-Kit antibody

c-Kit Ligand antibody

CD117 antibody

Kit antibody

Kit Ligand antibody

KIT oncogene antibody

KIT proto oncogene receptor tyrosine kinase antibody

KIT_HUMAN antibody

Mast cell growth factor receptor antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of c-Kit on different lysates with Rabbit anti-c-Kit antibody (<a href="/products/ET1609-60" style="font-weight: bold;text-decoration: underline;">ET1609-60</a>) at 1/2,000 dilution.<br /><br />Lane 1: Saos-2 cell lysate<br />Lane 2: HL-60 cell lysate (low expression)<br />Lane 3: Jurkat cell lysate (low expression)<br /><br />Lysates/proteins at 15 µg/Lane.<br /><br />Predicted band size: 110 kDa<br />Observed band size: 110-140 kDa<br /><br />Exposure time: 2 minutes 18 seconds;<br /><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/ET1609-60" style="font-weight: bold;text-decoration: underline;">ET1609-60</a>) at 1/2,000 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.

    ☑ Relative expression (RE)

    Western blot analysis of c-Kit on different lysates with Rabbit anti-c-Kit antibody (ET1609-60) at 1/2,000 dilution.

    Lane 1: Saos-2 cell lysate
    Lane 2: HL-60 cell lysate (low expression)
    Lane 3: Jurkat cell lysate (low expression)

    Lysates/proteins at 15 µg/Lane.

    Predicted band size: 110 kDa
    Observed band size: 110-140 kDa

    Exposure time: 2 minutes 18 seconds;

    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 (ET1609-60) at 1/2,000 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.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of c-Kit on different lysates with Rabbit anti-c-Kit antibody (<a href="/products/ET1609-60" style="font-weight: bold;text-decoration: underline;">ET1609-60</a>) at 1/5,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-c-Kit KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 110 kDa<br />Observed band size: 140 kDa<br /><br />Exposure time: 2 minute 40 seconds; ECL: K1801;<br /><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/ET1609-60" style="font-weight: bold;text-decoration: underline;">ET1609-60</a>) at 1/5,000 dilution was used in primary antibody dilution (K1803) 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.

    ☑ Knockdown (KD)

    Western blot analysis of c-Kit on different lysates with Rabbit anti-c-Kit antibody (ET1609-60) at 1/5,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-c-Kit KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 110 kDa
    Observed band size: 140 kDa

    Exposure time: 2 minute 40 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 (ET1609-60) at 1/5,000 dilution was used in primary antibody dilution (K1803) 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 c-Kit on human brain tissue lysates with Rabbit anti-c-Kit antibody (<a href="/products/ET1609-60" style="font-weight: bold;text-decoration: underline;">ET1609-60</a>) at 1/500 dilution.<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 110 kDa<br />Observed band size: 110 kDa<br /><br />Exposure time: 2 minutes;<br /><br />6% 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/ET1609-60" style="font-weight: bold;text-decoration: underline;">ET1609-60</a>) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:300,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of c-Kit on human brain tissue lysates with Rabbit anti-c-Kit antibody (ET1609-60) at 1/500 dilution.

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 110 kDa
    Observed band size: 110 kDa

    Exposure time: 2 minutes;

    6% 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 (ET1609-60) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human breast tissue with Rabbit anti-c-Kit antibody (<a href="/products/ET1609-60" style="font-weight: bold;text-decoration: underline;">ET1609-60</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1609-60" style="font-weight: bold;text-decoration: underline;">ET1609-60</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human breast tissue with Rabbit anti-c-Kit antibody (ET1609-60) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-60) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human gastrointestinal stromal tumor tissue with Rabbit anti-c-Kit antibody (<a href="/products/ET1609-60" style="font-weight: bold;text-decoration: underline;">ET1609-60</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1609-60" style="font-weight: bold;text-decoration: underline;">ET1609-60</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human gastrointestinal stromal tumor tissue with Rabbit anti-c-Kit antibody (ET1609-60) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-60) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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