Human CIS-1 Antibody

R&D Systems | Catalog # AF3194

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Western Blot, Immunocytochemistry, Simple Western

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all CIS-1 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human CIS-1
Leu11-Leu258
Accession # Q9NSE2

Specificity

Detects human CIS-1 in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human CIS-1 Antibody

Detection of Human CIS-1 antibody by Western Blot.

Detection of Human CIS-1 by Western Blot.

Western blot shows lysates of Nalm-6 human Pre-B acute lymphocytic leukemia cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human CIS-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3194) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for CIS-1 at approximately 35-40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

CIS‑1 in HDLM‑2 Human Cell Line.

CIS‑1 was detected in immersion fixed HDLM‑2 human Hodgkin’s lymphoma cell line using Goat Anti-Human CIS‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3194) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Detection of Human CIS‑1 by Simple WesternTM.

Simple Western lane view shows lysates of NK human natural killer lymphoma cell line, loaded at 0.2 mg/mL. Specific bands were detected for CIS‑1 at approximately 37 and 42 kDa (as indicated) using 10 µg/mL of Goat Anti-Human CIS‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3194). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Detection of CIS-1 by Western Blot

Detection of CIS-1 by Western Blot

CAGE regulates anticancer drug resistance and autophagic flux. (A) After 48 h of transfection with the indicated siRNA (each at 10 nM), immunoblot was performed. Representative blots of three independent experiments are shown. Ctrl. denotes control siRNA. (B) After 24 h of transfection, cells were treated with the indicated anticancer drug for 24 h. Quantification of band intensity was represented by the ratio of cleaved/full-length PARP normalized to actin. (C) Same as (A) except that immunofluorescence staining was performed. ***, p < 0.001. (D) After 48 h of transfection with the indicated construct, immunoblot was performed. Representative blots of three independent experiments are shown. (E) After 24 h of transfection, the cells were treated with the indicated anticancer drug (each at 1 μM) for 24 h. (F) Same as (D) except for the immunofluorescence staining. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34113619), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of CIS-1 by Western Blot

Detection of CIS-1 by Western Blot

CAGE expression is regulated by autophagy regulators. (A) AGS cells were treated with or without rapamycin for 24 h. Representative blots of three independent experiments are shown. (B) AGS cells were treated with or without rapamycin for 24 h, followed by immunofluorescence staining. ***, p < 0.001. (C) AGS cells were treated with or without rapamycin (5 μM) for 24 h, followed by treatment with celastrol (1 μM), Taxol (1 μM), or doxorubicin (1 μM) for 24 h. Representative blots of three independent experiments are shown. (D) AGSR cells were treated with or without chloroquine (CQ) (100 μM) for 24 h. Representative blots of three independent experiments are shown. (E) AGSR cells were treated with or without CQ (100 μM) for 24 h, followed by treatment with the indicated anticancer drug for 24 h. Representative blots of three independent experiments are shown. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34113619), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of CIS-1 by Western Blot

Detection of CIS-1 by Western Blot

Soluble factors regulate autophagic flux. (A) Culture medium of AGSR cells were added to AGS cells for 24 h. Representative blots of three independent experiments are shown. (B) Same as (A) except that the number of LC3 puncta was determined. ***, p < 0.001. (C) Cytokine array analysis was performed. (D) Matrigel plug assays using culture medium were performed. (E) Immunoblot was performed. Representative blots of three independent experiments are shown. (F) After 48 h of transfection with the indicated siRNA (each at 10 nM), culture medium was added to AGS cells for 24 h. Representative blots of three independent experiments are shown. (G) Culture medium of the indicated cancer cells was added to the CAGE CRISPR–Cas9 cell line for 24 h. Representative blots of three independent experiments are shown. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34113619), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of CIS-1 by Western Blot

Detection of CIS-1 by Western Blot

CAGE regulates tumorigenic potential in vivo. (a) The indicated cancer cells (each at 1 × 106) were injected into the dorsal flanks of athymic nude mice. Tumor volumes were measured as described. Significance determined by one-way ANOVA.***, p < 0.001. Each experimental group consisted of five athymic nude mice. (b) Tumor tissue lysates were subjected to immunoblot. Representative blots of three independent experiments were shown. The uncropped blots are shown in Supplementary Materials. (c) The indicated cancer cells (each at 1 × 106) were injected into the dorsal flanks of nude mice. Each experimental group consisted of five nude mice. Data are presented as a mean ± SEM. Tumor volumes were measured as described. Significance determined by one-way ANOVA. *, p < 0.05; ***, p < 0.001. (d) Tumor tissue lysates were subjected to immunoblot and immunohistochemical staining. Representative images of three independent experiments were shown. Data are presented as mean ± SEM. Significance determined by one-way ANOVA. *, p < 0.05; **, p < 0.01; ***, p < 0.001. The uncropped blots are shown in Supplementary Materials. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37735252), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of CIS-1 by Western Blot

Detection of CIS-1 by Western Blot

MiR-302b-5p regulates autophagic flux and anticancer drug resistance. (A) After 48 h of transfection with the indicated inhibitor (each at 10 nM), immunoblot and immunoprecipitation were performed. Representative blots of three independent experiments are shown. (B) Same as (A) except that immunofluorescence staining was performed. **, p < 0.01. (C) qRT-PCR was performed. **, p < 0.01. Average values of three independent experiments are shown. (D) AGS cells were transfected with the indicated inhibitor (each at 10 nM). The following day, cells were then treated with or without celastrol (1 μM), Taxol (1 μM), or doxorubicin (1 μM) for 24 h. Representative blots of three independent experiments are shown. (E) After 48 h of transfection with the indicated mimic (each at 10 nM), immunoblot and immunoprecipitation were performed. Representative blots of three independent experiments are shown. (F) Same as (E) except that the number of LC3 puncta was determined. **, p < 0.01. (G) qRT-PCR analysis was performed. ***, p < 0.001. Average values of three independent experiments are shown. (H) AGSR cells were transfected with the indicated mimic (each at 10 nM). The next day, cells were then treated with or without celastrol (1 μM), Taxol (1 μM), or doxorubicin (1 μM) for 24 h. Representative blots of three independent experiments are shown. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34113619), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of CIS-1 by Western Blot

Detection of CIS-1 by Western Blot

Soluble factors regulate autophagic flux. (A) Culture medium of AGSR cells were added to AGS cells for 24 h. Representative blots of three independent experiments are shown. (B) Same as (A) except that the number of LC3 puncta was determined. ***, p < 0.001. (C) Cytokine array analysis was performed. (D) Matrigel plug assays using culture medium were performed. (E) Immunoblot was performed. Representative blots of three independent experiments are shown. (F) After 48 h of transfection with the indicated siRNA (each at 10 nM), culture medium was added to AGS cells for 24 h. Representative blots of three independent experiments are shown. (G) Culture medium of the indicated cancer cells was added to the CAGE CRISPR–Cas9 cell line for 24 h. Representative blots of three independent experiments are shown. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34113619), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of CIS-1 by Western Blot

Detection of CIS-1 by Western Blot

MiR-302b-5p regulates autophagic flux and anticancer drug resistance. (A) After 48 h of transfection with the indicated inhibitor (each at 10 nM), immunoblot and immunoprecipitation were performed. Representative blots of three independent experiments are shown. (B) Same as (A) except that immunofluorescence staining was performed. **, p < 0.01. (C) qRT-PCR was performed. **, p < 0.01. Average values of three independent experiments are shown. (D) AGS cells were transfected with the indicated inhibitor (each at 10 nM). The following day, cells were then treated with or without celastrol (1 μM), Taxol (1 μM), or doxorubicin (1 μM) for 24 h. Representative blots of three independent experiments are shown. (E) After 48 h of transfection with the indicated mimic (each at 10 nM), immunoblot and immunoprecipitation were performed. Representative blots of three independent experiments are shown. (F) Same as (E) except that the number of LC3 puncta was determined. **, p < 0.01. (G) qRT-PCR analysis was performed. ***, p < 0.001. Average values of three independent experiments are shown. (H) AGSR cells were transfected with the indicated mimic (each at 10 nM). The next day, cells were then treated with or without celastrol (1 μM), Taxol (1 μM), or doxorubicin (1 μM) for 24 h. Representative blots of three independent experiments are shown. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34113619), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of CIS-1 by Western Blot

Detection of CIS-1 by Western Blot

Anticancer drug resistance is correlated with enhanced autophagy and antiapoptotic effects. (A) Cell lysates were subjected to immunoblot and immunoprecipitation. Representative blots of three independent experiments are shown. Immunofluorescence staining of LC3 (B) and cancer-associated gene (CAGE) (C) was performed. ***, p < 0.001. (D) Representative electron micrograph of AGS cells and AGSR cells is shown. The black arrows indicate autolysosomes. The white arrows indicate multivesicular bodies. (E) After 48 h of treatment, immunoblot was performed. C-PARP denotes cleaved PARP. Representative blots of three independent experiments are shown. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34113619), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human CIS-1 Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed HDLM‑2 human Hodgkin's lymphoma cell line

Simple Western

10 µg/mL
Sample: NK human natural killer lymphoma cell line

Western Blot

1 µg/mL
Sample: Nalm‑6 human Pre‑B acute lymphocytic leukemia cell line

Reviewed Applications

Read 1 review rated 1 using AF3194 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: CIS-1

Cytokine Inducible SH2-containing protein (CIS-1) is a 29 kDa protein found in a variety of cell types. Mono or polyubiquitination generally results in a 37 or 45 kDa molecule. CIS-1 binds to phosphorylated cytokine receptors IL-3 R beta and EPO-R and blocks downstream activation of STAT5 via receptor internalization and ubiquitin‑mediated proteosomal degradation. Human CIS-1 is a 258 aa peptide that contains one SH2 domain (aa 82‑163) and one SOCS box (aa 218‑258). There are two known alternatively spliced variants with a 7- or 13-aa substitution for the 7 N-terminal amino acid residues. Over the region used as immunogen, human CIS-1 is 91% identical to the corresponding mouse and canine protein sequences.

Long Name

Cytokine Inducible SH2-containing Protein

Alternate Names

CISH

Entrez Gene IDs

1154 (Human); 12700 (Mouse); 83681 (Rat)

Gene Symbol

CISH

UniProt

Q9NSE2

Additional CIS-1 Products

Product Documents for Human CIS-1 Antibody

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Product Specific Notices for Human CIS-1 Antibody

For research use only

Citations for Human CIS-1 Antibody

Customer Reviews for Human CIS-1 Antibody (1)

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  • Human CIS-1 Antibody
    Name: Maud TUSSEAU
    Application: Simple Western
    Sample Tested: Hek 293T cell lysate
    Species: Human
    Verified Customer | Posted 08/06/2021
    HEK937T transfected with SOCS1 WT lysates 4 types of lysis : NP40/LAEMMLI/CHAPS/TRITON
    Human CIS-1 Antibody AF3194
    Bio-Techne Response
    This review was submitted through the legacy Novus Innovators Program, reflecting a new species or application tested on a primary antibody.

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