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AB289996

Anti-HC-II antibody [EPR25618-1] - BSA and Azide free

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Rabbit Recombinant Monoclonal HC-II antibody. Carrier free. Suitable for ICC/IF, IP, WB, Flow Cyt (Intra) and reacts with Human samples.

View Alternative Names

HCF2, SERPIND1, Heparin cofactor 2, Heparin cofactor II, Protease inhibitor leuserpin-2, Serpin D1, HC-II, HLS2

5 Images
Flow Cytometry (Intracellular) - Anti-HC-II antibody [EPR25618-1] - BSA and Azide free (AB289996)
  • Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-HC-II antibody [EPR25618-1] - BSA and Azide free (AB289996)

This data was developed using ab289971, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HepG2 (Human hepatocellular carcinoma epithelial cell) cells labelling HC-II with ab289971 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

Immunocytochemistry/ Immunofluorescence - Anti-HC-II antibody [EPR25618-1] - BSA and Azide free (AB289996)
  • ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-HC-II antibody [EPR25618-1] - BSA and Azide free (AB289996)

This data was developed using ab289971, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 100% Methanol-fixed, 0.1% TritonX-100 permebilized HepG2 cells labelling HC-II with ab289971 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HepG2 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

Immunoprecipitation - Anti-HC-II antibody [EPR25618-1] - BSA and Azide free (AB289996)
  • IP

Lab

Immunoprecipitation - Anti-HC-II antibody [EPR25618-1] - BSA and Azide free (AB289996)

This data was developed using ab289971, the same antibody clone in a different buffer formulation.

HC-II was immunoprecipitated from HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate 10 ug with ab289971 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab289971 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate 10 ug

Lane 2 : ab289971 IP in HepG2 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab289971 in HepG2 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 10 seconds

All lanes:

Immunoprecipitation - Anti-HC-II antibody [EPR25618-1] (<a href='/en-us/products/primary-antibodies/hc-ii-antibody-epr25618-1-ab289971'>ab289971</a>)

Predicted band size: 57 kDa

Observed band size: 66-75 kDa

false

Western blot - Anti-HC-II antibody [EPR25618-1] - BSA and Azide free (AB289996)
  • WB

Lab

Western blot - Anti-HC-II antibody [EPR25618-1] - BSA and Azide free (AB289996)

This data was developed using ab289971, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST

All these lysates are RIPA buffer.

Exposure time : 59 seconds

All lanes:

Western blot - Anti-HC-II antibody [EPR25618-1] (<a href='/en-us/products/primary-antibodies/hc-ii-antibody-epr25618-1-ab289971'>ab289971</a>) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellar carcinoma epithelial cell) transfected with scrambled siRNA control, whole cell lysate at 20 µg

Lane 2:

HepG2 transfected with HC-II siRNA, whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 57 kDa

Observed band size: 66-75 kDa

false

Western blot - Anti-HC-II antibody [EPR25618-1] - BSA and Azide free (AB289996)
  • WB

Lab

Western blot - Anti-HC-II antibody [EPR25618-1] - BSA and Azide free (AB289996)

This data was developed using ab289971, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Exposure time : 15 seconds

All lanes:

Western blot - Anti-HC-II antibody [EPR25618-1] (<a href='/en-us/products/primary-antibodies/hc-ii-antibody-epr25618-1-ab289971'>ab289971</a>) at 1/1000 dilution

Lane 1:

Human liver tissue lysate(RIPA) at 20 µg

Lane 2:

Human liver tissue lysate(HOT) at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 57 kDa

Observed band size: 66-75 kDa

false

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR25618-1

Isotype

IgG

Carrier free

Yes

Reacts with

Human

Applications

WB, ICC/IF, Flow Cyt (Intra), IP

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>", "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "", "IP-species-notes": "<p></p>", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "<p></p>" }, "Mouse": { "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "" }, "Rat": { "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "" } } }
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AB277402

Human HC-II ELISA Kit

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We recommend this product because it’s often used in the same experiment or related research.

We advise that you always check the datasheet to ensure it fits your experiments, or contact ourtechnical teamfor help.

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Product details

ab289996 is the carrier-free version of ab289971.

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Constituents: 100% PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

HC-II also known as heparin cofactor II is an enzyme that acts as a serine protease inhibitor. It mainly inhibits thrombin playing a significant role in the regulation of coagulation. HC-II has an approximate molecular mass of 65 kDa and is primarily expressed in the liver and circulates in plasma. The HC protein is similar to another anticoagulant protein antithrombin but is specifically activated by dermatan sulfate.
Biological function summary

Heparin cofactor II plays an essential role in maintaining hemostatic balance. It does not function as part of a larger complex but individually regulates thrombin activity. HC-II inhibits fibrin formation and platelet aggregation controlling thrombin’s role in the coagulation cascade. The activity of HC-II is important for keeping blood clot formation under control preventing excessive clot formation which can lead to vascular diseases.

Pathways

Heparin cofactor II works in the coagulation pathway where it closely interacts with thrombin. Dermatan sulfate enhances its inhibitory effects on thrombin distinct from the action of heparin on antithrombin. Its regulation of thrombin makes it an important component in the hemostatic pathway maintaining normal circulation by balancing blood clotting and bleeding.

Heparin cofactor II is associated with thrombotic disorders including venous thromboembolism. Deficiency or malfunctioning of HC-II can lead to an increased risk of thrombosis due to inadequate inhibition of thrombin. Besides HC-II is associated with atherosclerosis where its interaction with dermatan sulfate could influence the progression of vascular lesions. The protein's role in modulating thrombin activity further connects it to coagulation factor V which can also impact thrombotic disease processes.
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

Thrombin inhibitor activated by the glycosaminoglycans, heparin or dermatan sulfate. In the presence of the latter, HC-II becomes the predominant thrombin inhibitor in place of antithrombin III (AT-III). Also inhibits chymotrypsin, but in a glycosaminoglycan-independent manner.. Peptides at the N-terminal of HC-II have chemotactic activity for both monocytes and neutrophils.. Isoform 2. Shows negligible inhibition, in vitro, of thrombin and tPA and no inhibition of factor Xa, in vitro.
See full target information SERPIND1
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions
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Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com