Anti-c-Kit Antibody Picoband™

Product Name

Anti-c-Kit Antibody Picoband™

View all CD117/c-kit Antibodies

SKU/Catalog Number

PB9258

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-c-Kit Antibody Picoband™ catalog # PB9258. Tested in Flow Cytometry, IHC, ICC, WB applications. This antibody reacts with Human.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-c-Kit Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9258)

Host

Rabbit

Contents

Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human c-Kit recombinant protein (Position: Q26-S285). Human c-Kit shares 66% amino acid (aa) sequence identity with mouse c-Kit.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins

Reactive Species

PB9258 is reactive to KIT in Human

Applications

PB9258 is guaranteed for Flow Cytometry, IHC, ICC, WB Boster Guarantee

Observed Molecular Weight

109 kDa

Calculated molecular weight

109.865kDa

Background of CD117/c-kit

SCFR (Mast/stem cell growth factor receptor), also known as KIT ONCOGENE or CD117, is a protein that in humans is encoded by the KIT gene. KIT was first described as the cellular homolog of the feline sarcoma viral oncogene v-kit. The KIT gene is mapped on 4q12. Kit was expressed on the surface of germ cells up to the pachytene stage. Signaling from the KIT receptor tyrosine kinase is essential for primordial germ cell growth both in vivo and in vitro. Determination of the KIT effectors acting in primordial germ cells has been hampered by the lack of effective methods to manipulate easily gene expression in these cells.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, By Heat
Immunohistochemistry (Frozen Section), 0.5-1μg/ml
Immunocytochemistry, 0.5-1μg/ml
Western blot, 0.1-0.5μg/ml
Flow Cytometry, 1-3μg/1x10⁶cells

Validation Images & Assay Conditions

Click image to see more details

Figure 1. Western blot analysis of C-Kit using anti-C-Kit antibody (PB9258).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours.
Lane 1: Recombinan Human C-Kit Protein 0.5ng
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-C-Kit antigen affinity purified polyclonal antibody (Catalog # PB9258) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for C-Kit at approximately 49KD. The expected band size for C-Kit is at 49KD.

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Figure 2. Western blot analysis of C-Kit using anti-C-Kit antibody (PB9258).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours.
Lane 1: HEPG2 Whole Cell Lysate
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-C-Kit antigen affinity purified polyclonal antibody (Catalog # PB9258) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for C-Kit at approximately 109KD. The expected band size for C-Kit is at 109KD.

Click image to see more details

Figure 3. IHC analysis of C-Kit using anti-C-Kit antibody (PB9258).
C-Kit was detected in paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-C-Kit Antibody (PB9258) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

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Figure 4. Flow Cytometry analysis of K562 cells using anti-C-Kit antibody (PB9258).
Overlay histogram showing K562 cells stained with PB9258 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-C-Kit Antibody (PB9258,1μg/1x106 cells) for 30 min at 20°C. DyLight488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For KIT (Source: Uniprot.org, NCBI)

Alternative Names

CD117 antigen; CD117; ckit; c-kit; EC 2.7.10; EC 2.7.10.1; KIT; PBT; piebald trait; Proto-oncogene c-Kit; proto-oncogene tyrosine-protein kinase Kit; SCF R; SCFR; SCFRmast/stem cell growth factor receptor; soluble KIT variant 1; Tyrosine-protein kinase Kit; v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog; v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene-like protein KIT C-Kit, CD117, MASTC, PBT, SCFR KIT proto-oncogene, receptor tyrosine kinase mast/stem cell growth factor receptor Kit|c-Kit protooncogene|p145 c-kit|piebald trait protein|proto-oncogene c-Kit|proto-oncogene tyrosine-protein kinase Kit|soluble KIT variant 1|tyrosine-protein kinase Kit|v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog|v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene-like protein

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