HA751827

CD45 Recombinant Rabbit Monoclonal Antibody [JE03-05]

cat.: HA751827

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human
Applications:	WB, IHC-P, IF-Tissue, IF-Cell, FC(Intra)
Clonality:	Monoclonal
Clone number:	JE03-05

Form:	Liquid
Storage condition:	Store at 2-8℃. Avoid freeze.
Storage buffer:	1*PBS (pH7.4).
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 147 kDa.
Isotype:	IgG
Immunogen:	Recombinant protein within Human CD45 aa 1,210-1,306 / 1,306.
Positive control:	Jurkat (Human T-lymphoblastic cells) cell lysate, Raji (Human Burkitt's lymphoma cell) cell lysate, THP-1 (Human acute monoblastic leukemia cell) cell lysate, human colon cancer tissue, human tonsil tissue, human spleen tissue, Jurkat (Human T-lymphoblastic cells).
Subcellular location:	Cell junction, Cell membrane, Membrane
Uniprot #:	SwissProt: P08575 Human
Alternative names:	B220 CD 45 CD45 CD45 antigen CD45R GP180 L-CA LCA Leukocyte common antigen loc Ly-5 LY5 Ly5, homolog of Lyt-4 OTTHUMP00000033813 OTTHUMP00000033816 OTTHUMP00000033817 OTTHUMP00000038574 Protein tyrosine phosphatase receptor type c polypeptide Protein tyrosine phosphatase, receptor type C protein tyrosine phosphatase, receptor type, C Protein tyrosine phosphatase, receptor type, c polypeptide Ptprc PTPRC_HUMAN Receptor-type tyrosine-protein phosphatase C T200 T200 glycoprotein T200 leukocyte common antigen

Images

Fig1: This data was developed using HA751827, the same antibody clone in a different buffer formulation.
Western blot analysis of CD45 on different lysates with Rabbit anti-CD45 antibody (HA751827) at 1/2,000 dilution.

Lane 1: Jurkat (Human T-lymphoblastic cells) cell lysate
Lane 2: 293T (Human embryonic kidney cell) cell lysate
Lane 3: Raji (Human Burkitt's lymphoma cell) cell lysate
Lane 4: MCF7 (Human breast cancer cell) cell lysate
Lane 5: THP-1 (Human acute monoblastic leukemia cell) cell lysate

Lysates/proteins at 15 µg/Lane.
Exposure time: 25 seconds; ECL: K1801

293T is a negative control (PMID: 19593443).
MCF7 is a negative control (PMID: 30678037).

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA751827, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 147 kDa
Observed band size: 250 kDa

Fig2: This data was developed using HA751827, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-CD45 antibody (HA751827) at 1/1,000 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 (HA751827) at 1/1,000 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.

	Fig3: This data was developed using HA751827, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-CD45 antibody (HA751827) at 1/1,000 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 (HA751827) at 1/1,000 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.
	Fig4: This data was developed using HA751827, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-CD45 antibody (HA751827) at 1/1,000 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 (HA751827) at 1/1,000 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.
	Fig5: This data was developed using HA751827, the same antibody clone in a different buffer formulation. Immunofluorescence analysis of paraffin-embedded human tonsil tissue labeling CD45 with Rabbit anti-CD45 antibody (HA751827) at 1/500 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA751827, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

Fig6: This data was developed using HA751827, the same antibody clone in a different buffer formulation.
Application: Immunocytochemistry (IF-cell)

Species: Human
Sample: Jurkat (Human T-lymphoblastic cells) / 293T (Human embryonic kidney cell)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: HA751827, 1/500, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

Negative control: 293T (PMID: 19593443).

Fig7: This data was developed using HA751827, the same antibody clone in a different buffer formulation.
Application: Flow Cytometry (Intra)

Species: Human
Sample: 293T (Human embryonic kidney cell) (left, negative) / Jurkat (Human T-lymphoblastic cells) (right)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Tween-20, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.

Primary antibody: HA751827 (1/5,000, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.

Negative control: 293T (PMID: 19593443).

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.