ET1602-22

CCR7 Recombinant Rabbit Monoclonal Antibody [SR36-04]

cat.: ET1602-22

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, IP
Clonality:	Monoclonal
Clone number:	SR36-04

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 43 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human CCR7 aa 13-62 / 378.
Positive control:	HDLM-2 cell lysate, Jurkat cell lysate, Daudi cell lysate, MCF7 cell lysate, mouse spleen tissue lysate, mouse pancreas tissue lysate, rat spleen tissue lysate, Raji cell lysate, EL4 cell lysate, RAW264.7 cell lysate, C6 cell lysate, Daudi, RAW264.7, C6, human tonsil tissue, human spleen tissue.
Subcellular location:	Cell membrane.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P IP	1:2,000-1:5,000 1:200-1:500 1:200 1:200 1-2μg/sample
Uniprot #:	SwissProt: P32248 Human \| P47774 Mouse Unigene: 229736 Rat
Alternative names:	BLR 2 BLR2 C C chemokine receptor type 7 C C CKR 7 C-C chemokine receptor type 7 C-C CKR-7 CC chemokine receptor 7 CC chemokine receptor type 7 CC CKR 7 CC-CKR-7 CCCKR7 CCR 7 CCR-7 Ccr7 CCR7_HUMAN CD 197 CD197 CD197 antigen CDw197 Chemokine (C C motif) receptor 7 Chemokine (C C) receptor 7 Chemokine C C motif receptor 7 Chemokine C C receptor 7 Chemokine receptor 7 Chemokine receptor 7-like protein CMKBR7 EBI 1 EBI1 Ebi1h EBV Induced G Protein Coupled Receptor 1 EBV-induced G-protein coupled receptor 1 Epstein Barr virus induced G protein coupled receptor Epstein Barr virus induced gene 1 Epstein-Barr virus-induced G-protein coupled receptor 1 EVI 1 EVI1 Lymphocyte Specific G Protein Coupled Peptide Receptor MGC108519 MIP 3 beta receptor MIP-3 beta receptor MIP3 Beta Receptor

Images

Fig1: All lanes: Western blot analysis of CCR7 with anti-CCR7 antibody (ET1602-22) at 1:5,000 dilution.
Lane 1/2: Wild-type Hela whole cell lysate (10 µg).
Lane 3/4: CCR7 fragment 1 knockdown Hela whole cell lysate (10 µg).
Lane 5/6: CCR7 fragment 2 knockdown Hela whole cell lysate (10 µg).

ET1602-22 was shown to specifically react with CCR7 in wild-type Hela cells. Weakened was observed when CCR7 knockdown sample was tested. Wild-type and CCR7 knockdown samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1602-22, 1/500) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of CCR7 on different lysates with Rabbit anti-CCR7 antibody (ET1602-22) at 1/5,000 dilution.

Lane 1: HDLM-2 cell lysate
Lane 2: Jurkat cell lysate
Lane 3: Daudi cell lysate
Lane 4: MCF7 cell lysate
Lane 5: Mouse spleen tissue lysate
Lane 6: Mouse pancreas tissue lysate (low expression)
Lane 7: Rat spleen tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 43 kDa
Observed band size: 43 kDa

Exposure time: 24 seconds;

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 (ET1602-22) at 1/5,000 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.

Fig3: Western blot analysis of CCR7 on different lysates with Rabbit anti-CCR7 antibody (ET1602-22) at 1/2,000 dilution.

Lane 1: Raji cell lysate
Lane 2: Daudi cell lysate
Lane 3: EL4 cell lysate
Lane 4: RAW264.7 cell lysate
Lane 5: C6 cell lysate
Lane 6: Rat spleen tissue lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 43 kDa
Observed band size: 43 kDa

Exposure time: 10 seconds; 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 (ET1602-22) 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.

Fig4: Immunocytochemistry analysis of Daudi cells labeling CCR7 with Rabbit anti-CCR7 antibody (ET1602-22) at 1/250 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS 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-CCR7 antibody (ET1602-22) at 1/250 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.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

	Fig5: Immunocytochemistry analysis of RAW264.7 cells labeling CCR7 with Rabbit anti-CCR7 antibody (ET1602-22) at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS 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-CCR7 antibody (ET1602-22) at 1/500 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. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
	Fig6: Immunocytochemistry analysis of C6 cells labeling CCR7 with Rabbit anti-CCR7 antibody (ET1602-22) at 1/200 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS 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-CCR7 antibody (ET1602-22) at 1/200 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. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
	Fig7: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-CCR7 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1602-22, 1/200) for 30 minutes 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.

Fig8: Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-CCR7 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1602-22, 1/200) for 30 minutes 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.

Fig9: CCR7 was immunoprecipitated from 0.2 mg Daudi cell lysate with ET1602-22 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using ET1602-22 at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: Daudi cell lysate (input)
Lane 2: ET1602-22 IP in Daudi cell lysate
Lane 3: Rabbit IgG instead of ET1602-22 in Daudi cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 8 seconds; ECL: K1802

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