HAVCR1 antibody (C-Term)

Catalog No. ABIN3044226

This anti-HAVCR1 antibody is a Rabbit Polyclonal antibody detecting HAVCR1 in WB, IHC, ICC and IF. Suitable for Human. Independently validated for use in Immunohistochemistry.

Quick Overview for HAVCR1 antibody (C-Term) (ABIN3044226)

Target: HAVCR1 (Hepatitis A Virus Cellular Receptor 1 (HAVCR1))

Reactivity: Human

Host: Rabbit

Clonality: Polyclonal

Conjugate: This HAVCR1 antibody is un-conjugated

Application: Western Blotting (WB), Immunohistochemistry (IHC), Immunocytochemistry (ICC), Immunofluorescence (IF)

Product Details anti-HAVCR1 Antibody

Binding Specificity: AA 332-348, C-Term

Purpose: Anti-TIM 1/HAVCR1 Antibody Picoband®

Sequence: IKALQNAVEK EVQAEDN

Cross-Reactivity (Details): No cross-reactivity with other proteins

Characteristics: Anti-TIM 1/HAVCR1 Antibody (ABIN3044226). Tested in IF, IHC, ICC, WB applications. This antibody reacts with Human. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.

Purification: Immunogen affinity purified.

Immunogen: A synthetic peptide corresponding to a sequence at the C-terminus of human TIM 1.

Isotype: IgG

Application Details

Application Notes: Western blot, 0.1-0.5 μg/mL, Human
Immunohistochemistry (Paraffin-embedded Section), 0.5-1 μg/mL, Human
Immunocytochemistry , 0.5-1 μg/mL, Human, -
Immunocytochemistry/Immunofluorescence, 5 μg/mL, Human
1. de Souza, A. J., Oriss, T. B., O'Malley, K. J., Ray, A., Kane, L. P. T cell Ig and mucin 1 (TIM-1) is expressed on in vivo-activated T cells and provides a costimulatory signal for T cell activation. Proc. Nat. Acad. Sci. 102: 17113-17118, 2005. 2. Feigelstock, D., Thompson, P., Mattoo, P., Zhang, Y., Kaplan, G. G. The human homolog of HAVcr-1 codes for a hepatitis A virus cellular receptor. J. Virol. 72: 6621-6628, 1998.

Comment:

Antibody can be supported by chemiluminescence kit ABIN921124 in WB, supported by ABIN921231 in IHC(P) and ICC.

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #300028 (Immunohistochemistry)

by: Human Protein Atlas

No.: #300028

Date: 12/13/2016

Antigen: HAVCR1

Method validated: Immunohistochemistry

Positive Control: kidney, colon/rectum

Notes: Passed. ABIN3044226 staining is consistent with RNA-seq data and published literature.

Validation Images

Staining of human rectum section in a tissue microarray with ABIN3044226. See the protocol for detailed information.

Full Methods

Primary Antibody: ABIN3044226

Secondary Antibody: HRP polymer (ThermoFisher Scientific, TL-125-PH)

Full Protocol:

• Sample preparation:
• • Fixation and TMA preparation according to PMID: 22688270.
  • Cut 4µm TMA sections using a waterfall microtome (ThermoFisher Scientific, HM355S).
  • Dry sections ON at RT and then back them for 12-24h at 50°C.
• Deparaffinization and rehydration in xylene and graded alcohol series:
• • xylene twice for 3min.
  • 100% ethanol for 1min.
  • 95% ethanol for 1min.
  • 0.3% H₂O₂ in 95% ethanol for 5min to block endogenous peroxidase.
  • 70% ethanol for 1min.
  • 50% ethanol for 1min.
  • distilled H₂O for 2min.
• Antigen Retrieval by Heat Induced Epitope Retrieval (HIER):
• • Boil the sections in Lab Vision Citrate buffer pH6.0 (ThermoFisher Scientific, TA-250-PM1X) for 4min at 125°C in a decloaking chamber (Biocare Medical).
  • Allow slides to cool down to 90°C in the decloaking chamber.
• Immunostaining in a Lab Vision Autostainer 480 (ThermorFisher Scientific) at RT with 300µl of reagents for each step:
• • Rinse sections in Lab Vision wash buffer with extra tween added to a final concentration of 0.2% (ThermoFisher Scientific, TA-999-TT and TA-125-TW) wash buffer.
  • Block sections with Lab Vision Ultra V-Block (ThermoFisher Scientific, TA-125-UB) for 5min.
  • Rinse sections 2x in wash buffer.
  • Incubate sections with primary rabbit anti-HAVCR1 antibody (antibodies-online, ABIN3044226) diluted 1:2000 for 30min.
  • Rinse sections 3x in wash buffer.
  • Incubate sections with labeled HRP polymer (ThermoFisher Scientific, TL-125-PH) for 30min.
  • Rinse sections 2x in wash buffer.
  • Develop in DAB Quanto solution (ThermoFisher Scientific, TL-125-QHDX) for 5min.
  • Rinse sections in distilled H₂O.
• Washing, counterstaining, and coverslipping in Autostainer XL (Leica Biosystems, ST5010) at RT:
• • Counterstaining in Mayer’s hematoxylin plus (HistoLab, 01820) for 7.5min.
  • Rinse sections in tap water for 5min.
  • Rinse sections in lithium carbonate water diluted 1:5 for 5min.
  • Dehydration in graded ethanol series and Neo-Clear (Merck Millipore, 109843) according to the manufacturer recommendations.
  • Automated mounting (Leica Biosystems, CV5030) of coverslip with Pertex (Histolab, 00871).
• Microscopy:
• • Image acquisition on a Leica Aperio AT2 and Aperio Scanscope AT at 20x magnification.

Experimental Notes: Based on published literature, HAVCR1 is widely expressed with highest expected levels in kidney and testis. ABIN3044226 stains kidney and colon/rectum where the protein should be enhanced according to RNA-seq data. Staining in kidney is not distinct but the staining is better in colon/rectum.

Handling

Format: Lyophilized

Reconstitution: Add 0.2 mL of distilled water will yield a concentration of 500 μg/mL.

Concentration: 500 μg/mL

Buffer: Each vial contains 5 mg BSA, 0.9 mg NaCl, 0.2 mg Na2HPO4, 0.05 mg Thimerosal, 0.05 mg Sodium azide.

Preservative: Thimerosal (Merthiolate), Sodium azide

Precaution of Use: This product contains Thimerosal (Merthiolate) and Sodium azide: POISONOUS AND HAZARDOUS SUBSTANCES which should be handled by trained staff only.

Handling Advice: Avoid repeated freezing and thawing.

Storage: 4 °C,-20 °C

Storage Comment: 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.

Expiry Date: 12 months

Target Details for HAVCR1

Target: HAVCR1 (Hepatitis A Virus Cellular Receptor 1 (HAVCR1))

Alternative Name: HAVCR1

Background:

Synonyms: Hepatitis A virus cellular receptor 1,HAVcr-1,Kidney injury molecule 1,KIM-1,T-cell immunoglobulin and mucin domain-containing protein 1,TIMD-1,T-cell immunoglobulin mucin receptor 1,TIM,TIM-1,T-cell membrane protein 1,HAVCR1,KIM1, TIM1, TIMD1,

Tissue Specificity: Widely expressed, with highest levels in kidney and testis. Expressed by activated CD4+ T-cells during the development of helper T-cells responses. .

Background: KIM1 (KIDNEY INJURY MOLECULE 1), also known as HAVCR1, HAVCR or TIM1, is a protein that in humans is encoded by the KIM1 gene. The KIM1 gene is mapped to 5q33.3. Biochemical, mutational, and cell adhesion analyses confirm that Tim1 is capable of homophilic Tim-Tim interactions. The features identified in murine KIM1 is conserved in human KIM1. The KIM1 protein is indeed a receptor for the virus through the infection of canine osteogenic sarcoma cells expressing HAVCR1 with HAV. Using a monoclonal antibody to mouse Tim1, Tim1 is expressed after activation of naive T cells and on T cells differentiated in Th2-polarizing conditions. Ectopic expression of KIM1 during mouse T-cell differentiation leads to production of the Th2-type cytokine Il4, but not the Th1-type cytokine Ifng. KIM1-expressing epithelial cells internalized apoptotic bodies, and Kim1 is ly responsible for phagocytosis in cultured primary rat tubule epithelial cells and in porcine and canine epithelial cell lines.

Sequence Similarities: Belongs to the immunoglobulin superfamily. TIM family.

Molecular Weight: 90-100 kDa

UniProt: Q96D42