710523

antibody from Invitrogen Antibodies

Targeting: KIT C-Kit, CD117, PBT, SCFR

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

710523

Provider

Invitrogen Antibodies

Product name

c-Kit Recombinant Polyclonal Antibody (2HCLC)

Antibody type

Polyclonal

Antigen

Synthetic peptide

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

2HCLC

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of c-Kit was performed by loading 20 µg of HeLa (lane1), K562 (lane2) and Jurkat (lane3) cell lysates using Novex®NuPAGE®4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Proteins were transferred to a PVDF membrane and blocked with 5 % skim milk for 1 hour at room temperature. c-Kit was detected at ~100 kDa using c-Kit Recombinant Rabbit Polyclonal Antibody (Product # 710523) at 0.5-1 µg/mL in 2.5 % skim milk at 4°C overnight on a rocking platform. Goat anti-Rabbit IgG-HRP Secondary Antibody (Product # G-21234) at 1:5000 dilution was used and chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of c-Kit in whole cell extracts from HeLa, K562, and Jurkat cells (lanes 1-3 respectively) using a c-Kit Recombinant Rabbit Polyclonal Antibody (Product # 710523) at a dilution of 1 µg/mL. Detection was performed using an HRP-conjugated Goat anti-Rabbit secondary antibody followed by chemiluminescence (ECL). Results show a band at ~80kDa.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of C- Kit (CD117) was done on 70% confluent log phase K562 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes; permeabilized with 0.25% Triton X-100 for 10 minutes followed by blocking with 5% BSA for 1 hour at room temperature. The cells were incubated with C- Kit (CD117) Recombinant Rabbit Polyclonal Antibody (Product # 710523) at 1 µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor 488 Goat anti-Rabbit IgG Secondary Antibody (Product # A-11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade Gold Antifade Mountant with DAPI (Product # S36938). Panel c is a merged image showing cytoplasmic localization of C- Kit (CD117). The images were captured at 20X magnification.