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Hck Recombinant Protein Antigen

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Summary
Reactivity HuSpecies Glossary
Applications AC

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Hck Recombinant Protein Antigen Summary

Description
A recombinant protein antigen with a N-terminal His6-ABP tag corresponding to human Hck.

Source: E.coli

Amino Acid Sequence: STFSTLQELVDHYKKGNDGLCQKLSVPCMSSKPQKPWEK
Source
E. coli
Protein/Peptide Type
Recombinant Protein Antigen
Gene
HCK
Purity
>80% by SDS-PAGE and Coomassie blue staining

Applications/Dilutions

Dilutions
  • Antibody Competition 10 - 100 molar excess
Application Notes
This peptide is useful as a blocking peptide for NBP2-57211 The protein was purified by IMAC chromatography and the expected concentration is greater than 0.5 mg/ml. This protein is produced on demand, estimated shipping date is 4 weeks after the order is placed. For further blocking peptide related information and a protocol, click here.
Theoretical MW
22 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

Storage
Store at -20C. Avoid freeze-thaw cycles.
Buffer
PBS and 1M Urea, pH 7.4.
Preservative
No Preservative
Purity
>80% by SDS-PAGE and Coomassie blue staining

Notes

This product is intended to be used as a blocking antigen for antibody competition assays. Any other use of this antigen is done at the risk of the user. The use of this product for commercial production is strictly prohibited. Please contact technical support if you have any questions.

Alternate Names for Hck Recombinant Protein Antigen

  • Bmk
  • EC 2.7.10
  • EC 2.7.10.2
  • Hck
  • Hctk
  • hemopoietic cell kinaseJTK9
  • JTK9
  • p59-HCK/p60-HCK
  • tyrosine-protein kinase HCK

Background

The 60-kilodalton protein encoded by HCK resembles the product of the proto-oncogene c-src and is therefore likely to be a peripheral membrane protein (1). Protein-tyrosine kinase Hck is primarily expressed in hematopoietic cells, particularly granulocytes. Protein-tyrosine kinases are implicated in the control of cell growth by virtue of their frequent appearance as products of retroviral oncogenes and as components of growth factor receptors (2). Tyrosine kinases of the Src family are regulated via their Src homology 2 (SH2) and SH3 domains. The Nef protein of human immunodeficiency virus-1 (HIV-1) has previously been shown to bind with high affinity and specificity in vitro to the SH3 domain of Hck, a Src family member expressed primarily in myeloid cells. Results provide direct evidence that SH3 engagement is sufficient to activate a Src family kinase in vivo and suggest that Hck may be activated by Nef in HIV-infected macrophages (3).

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. This product is guaranteed for 1 year from date of receipt.

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Src [p Tyr419]
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AF2685
Western blot shows lysates of MDA‑MB‑468 human breast cancer cell line and HepG2 human hepatocellular carcinoma cell line untreated (-) or treated (+) with 50 μM pervanadate (PV) for 10 minutes. PVDF membrane was probed with 1 µg/mL of Rabbit Anti-Human Phospho-Src (Y419) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2685), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-Src (Y419) at approximately 56 - 61 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Simple Western lane view shows lysates of MBA‑MB‑468 human breast cancer cell line untreated (-) or treated (+) with 50 µM Pervanadate (PV) for 10 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-Src (Y419) at approximately 64 kDa (as indicated) using 10 µg/mL of Rabbit Anti-Human Phospho-Src (Y419) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2685). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Src phosphorylated at Y419 was detected in immersion fixed paraffin-embedded sections of human colon using Rabbit Anti-Human Phospho-Src (Y419) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2685) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue). Specific labeling was localized to the cytoplasm of epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
(1 Review)
(6 Publications)
Species: Hu
Applications: IHC, Simple Western, WB
Lyn Antibody
[Unconjugated]
AF3206
Western blot shows lysates of U937 human histiocytic lymphoma cell line, PT18 mouse mast/basophil cell line, and Y3‑Ag rat myeloid cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse/Rat Lyn Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3206) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for Lyn at approximately 56 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Simple Western lane view shows lysates of Nalm‑6 human Pre-B acute lymphocytic leukemia cell line, THP‑1 human acute monocytic leukemia cell line, Raji human Burkitt's lymphoma cell line, and Ramos human Burkitt's lymphoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Lyn at approximately 58-60 kDa (as indicated) using 5 µg/mL of Goat Anti-Human/Mouse/Rat Lyn Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3206) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Lyn was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) treated with monensin using Goat Anti-Human/Mouse/Rat Lyn Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3206) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
(2 Publications)
Species: Hu, Mu, Rt
Applications: ICC, Simple Western, WB
Fgr Antibody
NBP1-85677
Immunohistochemistry-Paraffin: Fgr Antibody [NBP1-85677] - Staining of human cerebral cortex shows low expression as expected.
Immunohistochemistry-Paraffin: Fgr Antibody [NBP1-85677] - Analysis in human bone marrow and skeletal muscle tissues using NBP1-85677 antibody. Corresponding FGR RNA-seq data are presented for the same tissues.
Immunohistochemistry-Paraffin: Fgr Antibody [NBP1-85677] - Staining of human Bone marrow shows strong cytoplasmic and membranous positivity in hematopoietic cells.
Species: Hu
Applications: ICC/IF, IHC, IHC-P, WB
Fyn Antibody
(FYN-01) - BSA Free
NB500-517
Immunoprecipitation: Fyn Antibody (FYN-01) [NB500-517] - Immunoprecipitation of Fyn from the lysate of T cells isolated from fresh buffy coats. Western blot was immunostained with anti-Fyn (FYN-01). Lane 1: original lysate of T cells Lane 2-3: Immunoprecipitated material eluted from affinity sorbent (FYN-01 coupled to Sepharose beads). Lanes differ in amount of T cell lysate loaded on the immunosorbent.
Immunocytochemistry/Immunofluorescence: Fyn Antibody (FYN-01) [NB500-517] - Staining of Fyn in human primary fibroblasts using anti-Fyn (FYN-01; green). Actin cytoskeleton was decorated by phalloidin (red) and cell nuclei stained with DAPI (blue).
Immunocytochemistry/Immunofluorescence: Fyn Antibody (FYN-01) [NB500-517] - Staining of Fyn in human HeLa cell line using anti-Fyn (FYN-01; green). Actin cytoskeleton was decorated by phalloidin (red) and cell nuclei stained with DAPI (blue).
(1 Review)
(4 Publications)
Species: Hu, Mu
Applications: ICC/IF, IHC, IHC-P, IP, WB
Lck [p Tyr394]
Antibody (755103)
MAB7500
Lck phosphorylated at Y394 was detected in immersion fixed Jurkat human acute T cell leukemia cells treated with (upper panel) or without (lower panel) Pervanadate using Mouse Anti-Human Phospho-Lck (Y394) Monoclonal Antibody (Catalog # MAB7500) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces and cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Western blot shows lysates of Jurkat human acute T cell leukemia cell line and Ramos human Burkitt's lymphoma cell line untreated (-) or treated (+) with 1 mM Pervanadate (PV) for 5 minutes. PVDF membrane was probed with 0.1 µg/mL of Mouse Anti-Human Phospho-Lck (Y394) Monoclonal Antibody (Catalog # MAB7500) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Phospho-Lck (Y394) at approximately 56 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
(7 Publications)
Species: Hu
Applications: ICC, WB
TNF-alpha
[Unconjugated]
210-TA
1 µg/lane of Recombinant Human TNF-alpha was resolved by SDS-PAGE with silver staining, under reducing (R) conditions, showing a band at 17 kDa.
Recombinant Human TNF-alpha (Catalog # 210‑TA) induces cytotoxicity in the L-929 mouse fibroblast cell line in the presence of the metabolic inhibitor actinomycin D. The ED50 for this effect is 25‑100 pg/mL.
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(738 Publications)
Species: Hu
Applications: BA
SYK Antibody
NBP1-32945
Western Blot: SYK Antibody [NBP1-32945] - SYK antibody detects SYK protein by western blot analysis. Whole cell extracts (30 ug) was separated by 7.5% SDS-PAGE, and the membrane was blotted with SYK antibody at a dilution of 1:2000. HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
Western Blot: SYK Antibody [NBP1-32945] - SYK antibody detects SYK protein by western blot analysis. Whole cell extracts (30 ug) was separated by 7.5% SDS-PAGE, and the membrane was blotted with SYK antibody at a dilution of 1:2000. HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
Immunoprecipitation: SYK Antibody [NBP1-32945] - Immunoprecipitation of SYK protein from A431 whole cell extracts using 5 ug of SYK antibody. Western blot analysis was performed using SYK antibody.
Species: Hu, Mu, Rt
Applications: ICC/IF, IHC, IHC-P, IP, WB
PYK2/FAK2 [p
Tyr402] Antibody (592 ...
MAB6210
Simple Western lane view shows lysates of Raji human Burkitt's lymphoma cell line untreated (-) or treated (+) with 0.2 mg/mL Pervanadate (PV) for 30 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-PYK2/FAK2 (Y402) at approximately 113 kDa (as indicated) using 5 µg/mL of Mouse Anti-Human Phospho-PYK2/FAK2 (Y402) Monoclonal Antibody (Catalog # MAB6210). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Western blot shows lysates of Raji human Burkitt's lymphoma cell line and Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with 1 mM Pervanadate (PV) for 30 minutes and 10 µg/mL Mouse Anti-Human CD3 epsilon Monoclonal Antibody (Catalog # MAB100) for 15 minutes. PVDF membrane was probed with 0.5 µg/mL of Mouse Anti-Human Phospho-PYK2/FAK2 (Y402) Monoclonal Antibody (Catalog # MAB6210) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). Specific bands were detected for Phospho-PYK2/FAK2 (Y402) at approximately 105-115 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
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ERK2 Antibody
(OTI6E5)
NBP1-47842
Immunohistochemistry-Paraffin: ERK2 Antibody (6E5) [NBP1-47842] - Endometrium tissue within the normal limits using antiMAPK1mouse monoclonal antibody. (Heat-induced epitope retrieval by 10mM citric buffer, pH6.0, 100C for 10min,Dilution 1:50)
Immunoprecipitation: ERK2 Antibody (6E5) [NBP1-47842] - TrueMab monoclonal anti-ERK2 antibody (Negative control: IP without adding anti-ERK2 antibody). For each experiment, 500ul of DDK tagged ERK2 overexpression lysates (at 1:5 dilution with HEK293T lysate), 2ug of antibody.
Immunohistochemistry-Paraffin: ERK2 Antibody (6E5) [NBP1-47842] - Staining of paraffin-embedded Adenocarcinoma of colon tissue using anti-ERK2 mouse monoclonal antibody.
Species: Ca, Hu, Pm, Mu, Rt
Applications: IHC, IHC-P, IP, WB
c-Abl Antibody
[Unconjugated]
AF5414
Western blot shows lysates of Jurkat human acute T cell leukemia cell line, MCF‑7 human breast cancer cell line, and MDA‑MB‑453 human breast cancer cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human c-Abl Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5414) followed by HRP-conjugated Anti‑Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for c-Abl at approximately 135 - 145 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for c‑Abl at approximately 149 kDa (as indicated) using 10 µg/mL of Goat Anti-Human c‑Abl Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5414) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the12-230 kDa separation system.
(1 Publication)
Species: Hu
Applications: Simple Western, WB
IL-6 [HRP]
D6050
(693 Publications)
Species: Hu
Applications: ELISA
ERK1 Antibody (1E5)
NBP2-22203
Flow Cytometry: ERK1 Antibody (1E5) [NBP2-22203] - Flow cytometric analysis of Hela cells using ERK1 mouse mAb (blue) and negative control (red).
Immunocytochemistry/Immunofluorescence: ERK1 Antibody (1E5) [NBP2-22203] - Analysis of NIH/3T3 cells using ERK1 mouse mAb (green). Blue: DRAQ5 fluorescent DNA dye. Red: Actin filaments have been labeled with Alexa Fluor-555 phalloidin.
Immunohistochemistry-Paraffin: ERK1 Antibody (1E5) [NBP2-22203] - Analysis of paraffin-embedded bladder cancer tissues using ERK1 mouse mAb with DAB staining.
Species: Hu, Pm, Mu, Rt
Applications: ELISA, Flow, ICC/IF, IHC, IHC-P, WB
NOD2 Antibody
(2D9) - BSA Free
NB100-524
Flow Cytometry: NOD2 Antibody (2D9) [NB100-524] - An intracellular stain was performed on Jurkat cells with NOD2 (2D9) antibody NB100-524APC (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes.
Flow Cytometry: NOD2 Antibody (2D9) [NB100-524] - An intracellular stain was performed on Jurkat cells with NOD2 (2D9) antibody NB100-524PE (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes.Both antibodies were conjugated to phycoerythrin.
Immunohistochemistry-Frozen: NOD2 Antibody (2D9) [NB100-524] - Overlay of NOD2-DyLight 488 (green) with phase contrast of murine colon. Image from verified customer review.
(1 Review)
(24 Publications)
Species: Hu, Mu
Applications: Flow-IC, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IP, WB
HLA DQ/DR/DP
Antibody (HLA-Pan/296 ...
NBP2-79843
Protein Array: HLA DQ/DR/DP Antibody (HLA-Pan/2967R) - Azide and BSA Free [NBP2-79843] - Analysis of Protein Array containing more than 19,000 full-length human proteins using HLA DQ/DR/DP Antibody (HLA-Pan/2967R). Z- and S- Score: The Z-score represents the strength of a signal that a monoclonal antibody (MAb) (in combination with a fluorescently-tagged anti-IgG secondary antibody) produces when binding to a particular protein on the HuProt(TM) array. Z-scores are described in units of standard deviations (SD's) above the mean value of all signals generated on that array. If targets on HuProt(TM) are arranged in descending order of the Z-score, the S-score is the difference (also in units of SD's) between the Z-score. S-score therefore represents the relative target specificity of a MAb to its intended target. A MAb is considered to specific to its intended target, if the MAb has an S-score of at least 2.5.
Immunocytochemistry/Immunofluorescence: HLA DQ/DR/DP Antibody (HLA-Pan/2967R) - Azide and BSA Free [NBP2-79843] - Immunofluorescence staining of Raji cells using HLA DQ/DR/DP Antibody (HLA-Pan/2967R) followed by goat anti-rabbit IgG-CF488 (Green). Nuclei stained with RedDot (Red).
Western Blot: HLA DQ/DR/DP Antibody (HLA-Pan/2967R) - Azide and BSA Free [NBP2-79843] - Western Blot Analysis of Ramos cell lysate using HLA DQ/DR/DP Antibody (HLA-Pan/2967R).
Species: Hu
Applications: CyTOF-ready, ELISA, Flow, ICC/IF, IHC, IHC-P, PA, WB
CTLA-4
[Unconjugated]
7268-CT
Recombinant Human CTLA-4 Fc Chimera (Catalog # 7268-CT) inhibits IL-2 secretion by stimulated Jurkat human acute Tcell leukemia cells. The ED50 for this effect is 0.03-0.15 μg/mL whenstimulated with 1 μg/mL Recombinant Human B7‑1/CD80 Fc Chimera (Catalog # 140-B1).
(3 Publications)
Species: Hu
Applications: BA
AKT [p Ser473]
Antibody - Pan Specif
AF887
Western blot shows lysates of NIH‑3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 100 ng/mL Human PDGF (Catalog # 120‑HD) for 15 minutes and MCF‑7 human breast cancer cell line untreated or treated with 100 ng/mL Recombinant Human IGF-1 (Catalog # 291‑G1) for 15 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-Akt (S473) Pan Specific Antigen Affinity-purified Polyclonal Antibody (Catalog # AF887), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-Akt (S473) Pan Specific at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Simple Western lane view shows lysates of MCF‑7 human breast cancer cell line and A549 human lung carcinoma cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human IGF‑I (Catalog # 291-G1) for 15 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-Akt (S473) at approximately 60 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-Akt (S473) Pan Specific Antigen Affinity-purified Polyclonal Antibody (Catalog # AF887). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Serum-starved MCF‑7 human breast cancer cells were unstimulated (light orange filled histogram) or activated with 100 ng/mL Recombinant human IGF-1 (Catalog # 291-G1, dark orange filled histogram) for 15 minutes, then stained with Rabbit Anti-Human/Mouse/Rat Phospho‑Akt (S473) Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF887) or control antibody (Catalog # AB-105-C, open histogram), followed by Allophycocyanin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0111). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.
(28 Publications)
Species: Hu, Mu, Rt
Applications: CyTOF-ready, IHC, ICFlow, Simple Western, WB
Inorganic
Pyrophosphatase/PPA1
NBP1-31348
Immunocytochemistry/Immunofluorescence: Inorganic Pyrophosphatase/PPA1 Antibody [NBP1-31348] - Paraformaldehyde-fixed HeLa, using antibody at 1:200 dilution.
Immunohistochemistry-Paraffin: Inorganic Pyrophosphatase/PPA1 Antibody [NBP1-31348] - Paraffin-embedded OVCAR3 xenograft, using antibody at 1:500 dilution.
Western Blot: Inorganic Pyrophosphatase/PPA1 Antibody [NBP1-31348] - Sample (30 ug of whole cell lysate) A: H1299 B: Hela C: Hep G2 D: Molt-4 12% SDS PAGE; antibody diluted at 1:10000.
Species: Hu, Mu, Rt
Applications: ICC/IF, IHC, IHC-P, WB
Hck Recombinant
Protein Antigen
NBP2-57211PEP
Hck Recombinant Protein Antigen
Species: Hu
Applications: AC

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Bioinformatics

Gene Symbol HCK