Human CIS-1 Antibody

Catalog #: AF3194 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF3194
AF3194-SP

Detection of Human CIS-1 by Western Blot.

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Human CIS-1 Antibody Summary

Species Reactivity

Human

Specificity

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

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

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

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.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

Simple Western

10 µg/mL

NK human natural killer lymphoma cell line

Immunocytochemistry

5-15 µg/mL

Immersion fixed HDLM‑2 human Hodgkin's lymphoma cell line

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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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.

Immunocytochemistry

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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.

Simple Western

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Detection of Human CIS‑1 by Simple Western^TM. 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.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C

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.

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

Entrez Gene IDs

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

Alternate Names

CIS-1; CIS-1cytokine-inducible SH2-containing protein; CIScytokine-inducible inhibitor of signaling type 1B; CISH; cytokine inducible SH2-containing protein; G18Protein G18; SOCS; Suppressor of cytokine signaling