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ICAM3/CD50 Rabbit mAb (A25722)

Tested applications:WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP InhibitionReactivity:Human

Western blot analysis of various lysates using ICAM3/CD50 Rabbit mAb (A25722)at 1:1600 dilution incubated overnight at 4°C.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020)
Negative control (NC): Daudi
Exposure time: 1s.

ABclonal:Immunohistochemistry - ICAM3/CD50 Rabbit mAb (A25722)

Immunohistochemistry analysis of paraffin-embedded Human tonsil tissue using ICAM3/CD50 Rabbit mAb (A25722) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human spleen tissue using ICAM3/CD50 Rabbit mAb (A25722) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

ABclonal:Immunofluorescence - ICAM3/CD50 Rabbit mAb (A25722)

Confocal imaging of Jurkat cells using ICAM3/CD50 Rabbit mAb (A25722, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). Objective: 100x.

Confocal imaging of paraffin-embedded Human colon cancer tissue using ICAM3/CD50 Rabbit mAb (A25722, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). High pressure antigen retrieval performed with 0.01M Citrate Buffer (pH 6.0) prior to IF staining. Objective: 40x.

Confocal imaging of U-937 cells using ICAM3/CD50 Rabbit mAb (A25722, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). Objective: 100x.

ABclonal:Flow CytoMetry - ICAM3/CD50 Rabbit mAb (A25722)

Flow cytometry: 1X10^6 Daudi cells (negative control, left) and U-937 cells (right) were surface-stained with ICAM3/CD50 Rabbit mAb (A25722, 2 μg/mL, orange line), or Rabbit IgG isotype control (AC042, 2 μg/mL, blue line), followed by Alexa Fluor® 647 conjugated goat anti-rabbit pAb staining. Non-fluorescently stained cells were used as blank control (red line).

Flow cytometry: 1X10^6 U-937 cells were surface-stained with Rabbit IgG isotype control (AC042, 2 μg/mL, left) or ICAM3/CD50 Rabbit mAb (A25722, 2 μg/mL, right), followed by Alexa Fluor® 647 conjugated goat anti-rabbit pAb staining.

Overview

Product name	ICAM3/CD50 Rabbit mAb
Catalog No.	A25722
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG

Background

The protein encoded by this gene is a member of the intercellular adhesion molecule (ICAM) family. All ICAM proteins are type I transmembrane glycoproteins, contain 2-9 immunoglobulin-like C2-type domains, and bind to the leukocyte adhesion LFA-1 protein. This protein is constitutively and abundantly expressed by all leucocytes and may be the most important ligand for LFA-1 in the initiation of the immune response. It functions not only as an adhesion molecule, but also as a potent signalling molecule. Alternative splicing results in multiple transcript variants encoding different isoforms.

Immunogen information

Immunogen	Recombinant fusion protein containing a sequence corresponding to amino acids 30-114 of human ICAM3 (NP_002153.2).
Sequence	QEFLLRVEPQNPVLSAGGSLFVNCSTDCPSSEKIALETSLSKELVASGMGWAAFNLSNVTGNSRILCSVYCNGSQITGSSNITVY
Gene ID
Swiss Prot
Synonyms	CD50; CDW50; ICAM-R
Calculated MW	60kDa
Observed MW	140kDa

Applications

Reactivity	Human
Tested applications	WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP Inhibition
Recommended dilution	WB 1:1000 - 1:5000 IHC-P 1:200 - 1:2000 IF/ICC 1:200 - 1:800 FC 1:500 - 1:1000 ELISA Recommended starting concentration is 1 μg/mL. Please optimize the concentration based on your specific assay requirements.
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.05% proclin300, 0.05% BSA, 50% glycerol, pH7.3.
Key application	Western blotting Immunohistochemistry Immunofluorescence Flow Cytometry
Positive samples	RPMI 8226
Cellular location	Membrane, Single-pass type I membrane protein.