CD32 Recombinant Rabbit Monoclonal Antibody [JM10-70]
Catalog# ET1703-43
CD32 Recombinant Rabbit Monoclonal Antibody [JM10-70]
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WB
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IHC-P
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IF-Cell
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FC
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IP
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Human
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HA750382
不含抗保成分
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unconjugated
Overview
Product Name
CD32 Recombinant Rabbit Monoclonal Antibody [JM10-70]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within Human CD32 aa 23-274 / 310.
Species Reactivity
Human
Validated Applications
WB, IHC-P, IF-Cell, FC, IP
Molecular Weight
Predicted band size: 35 kDa
Positive Control
THP-1 cell lysate, K-562 cell lysate, human spleen tissue, K-562.
Conjugation
unconjugated
Clone Number
JM10-70
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:1,000-1:5,000
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IHC-P
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1:400
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IF-Cell
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1:100
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FC
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1:1,000
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IP
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1-2μg/sample
Target
Function
CD32 (also designated Fc gamma RII) is a low affinity receptor for the Fc fragment of aggregated IgG. CD32 is responsible for the clearance of immunocomplexes by macrophages and also plays an important role in the regulation of antibody production by B cells. IgG can noncooperatively bind either one or two highly glycosylated CD32 molecules, and this binding delivers a negative signal for B cells . CD32 exists as several isoforms that are produced by alternative splicing of three distinct genes, A, B, and C . These isoforms are designated FcgRIIA, FcgRIIB1, FcgRIIB3, and FcgRIIC . All isoforms are present on monocytes, placental trophoblasts and endothelial cells . In addition, the FcgRIIB forms are present on B lymphocytes, and the FcgRIIA and FcgRIIC forms are found on neutrophils.
Background References
1. Sips M et al. Fc receptor-mediated phagocytosis in tissues as a potent mechanism for preventive and therapeutic HIV vaccine strategies. Mucosal Immunol 9:1584-1595 (2016).
2. Karagiannis P et al. IgG4 subclass antibodies impair antitumor immunity in melanoma. J Clin Invest 123:1457-74 (2013).
Tissue Specificity
Found on monocytes, neutrophils and eosinophil platelets.
Post-translational Modification
Phosphorylated by SRC-type Tyr-kinases such as LYN, BLK, FYN, HCK and SYK.
Subcellular Location
Cell membrane.
UNIPROT
Synonyms
CD32 antibody
CD32A antibody
CD32B antibody
CD32C antibody
CDw32 antibody
Fc fragment of IgG low affinity IIa receptor antibody
Fc fragment of IgG low affinity IIa receptor for (CD32) antibody
Fc fragment of IgG low affinity IIb receptor antibody
Fc fragment of IgG low affinity IIb receptor for (CD32) antibody
Fc fragment of IgG low affinity IIc receptor antibody
ExpandImages
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☑ Cell treatment (CT)
Western blot analysis of CD32 on different lysates with Rabbit anti-CD32 antibody (ET1703-43) at 1/2,000 dilution.
Lane 1: THP-1 cell lysate
Lane 2: THP-1 cell lysate treated with deglycosylation
Lane 3: K-562 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 35 kDa
Observed band size: 40/31 kDa
Exposure time: 3 minutes; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1703-43) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-CD32 antibody (ET1703-43) at 1/400 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-43) at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunocytochemistry analysis of K-562 cells labeling CD32 with Rabbit anti-CD32 antibody (ET1703-43) at 1/100 dilution.
Cells were fixed in 80% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CD32 antibody (ET1703-43) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. -
Flow cytometric analysis of K-562 cells labeling CD32.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET1703-43, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
CD32 was immunoprecipitated from 0.2 mg THP-1 cell lysate with ET1703-43 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1703-43 at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: THP-1 cell lysate (input)
Lane 2: ET1703-43 IP in THP-1 cell lysate
Lane 3: Rabbit IgG instead of ET1703-43 in THP-1 cell lysate
Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 1 minute 7 seconds; ECL: K1801
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Products with the same target and pathway
CD32 Rabbit Polyclonal Antibody
Application: WB,IHC-P
Reactivity: Human,Mouse,Rat
Conjugate: unconjugated
