710001

antibody from Invitrogen Antibodies

Targeting: CCL5 D17S136E, MGC17164, RANTES, SCYA5, SISd, TCP228

Western blot (Data presented on provider website)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

710001

Provider

Invitrogen Antibodies

Product name

CCL5 (RANTES) Recombinant Superclonal™ Antibody (25HCLC)

Antibody type

Other

Antigen

Other

Description

This antibody is predicted to react with rat and human based on sequence homology. Recombinant rabbit Superclonal™ antibodies are unique offerings from Thermo Fisher Scientific. They are comprised of a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds - the sensitivity of polyclonal antibodies with the specificity of monoclonal antibodies - all delivered with the consistency only found in a recombinant antibody. While functionally the same as a polyclonal antibody - recognizing multiple epitope sites on the target and producing higher detection sensitivity for low abundance targets - a recombinant rabbit Superclonal™ antibody has a known mixture of light and heavy chains. The exact population can be produced in every lot, circumventing the biological variability typically associated with polyclonal antibody production. Note: Formerly called “Recombinant polyclonal antibody”, this product is now rebranded as “Recombinant Superclonal™ antibody”. The physical product and the performance remain unchanged.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Antibody clone number

25HCLC

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

Submitted references

PARP inhibitors promote stromal fibroblast activation by enhancing CCL5 autocrine signaling in ovarian cancer.

Li X, Fang T, Xu S, Jin P, Zhou D, Wang Z, Li H, Yang Z, Chen G, Zheng X, Xia Y, Wei X, Zhang Z, Yang X, Wang Y, Gao Q
NPJ precision oncology 2021 Jun 9;5(1):49

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Dysregulated oxalate metabolism is a driver and therapeutic target in atherosclerosis.

Liu Y, Zhao Y, Shukha Y, Lu H, Wang L, Liu Z, Liu C, Zhao Y, Wang H, Zhao G, Liang W, Fan Y, Chang L, Yurdagul A Jr, Pattillo CB, Orr AW, Aviram M, Wen B, Garcia-Barrio MT, Zhang J, Liu W, Sun D, Hayek T, Chen YE, Rom O
Cell reports 2021 Jul 27;36(4):109420

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DSE regulates the malignant characters of hepatocellular carcinoma cells by modulating CCL5/CCR1 axis.

Liao WC, Yen HR, Liao CK, Tseng TJ, Lan CT, Liu CH
American journal of cancer research 2019;9(2):347-362

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NADPH Oxidase 4 Regulates Inflammation in Ischemic Heart Failure: Role of Soluble Epoxide Hydrolase.

Stevenson MD, Canugovi C, Vendrov AE, Hayami T, Bowles DE, Krause KH, Madamanchi NR, Runge MS
Antioxidants & redox signaling 2019 Jul 1;31(1):39-58

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of CCL5/RANTES was performed using 70% confluent log phase HEK-293 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with CCL5/RANTES (25HCLC) Recombinant Rabbit Polyclonal Antibody (Product # 710001) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of CCL5/RANTES was performed using 70% confluent log phase HEK-293 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with CCL5/RANTES (25HCLC) Recombinant Rabbit Superclonal™ Antibody (Product # 710001) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of CCL5/RANTES showing staining in the cytoplasm of paraffin-embedded mouse lung tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a CCL5/RANTES Recombinant Rabbit Superclonal™ Antibody (Product # 710001) diluted in 3% BSA-PBS at a dilution of 1:20 for 1 hour at 37ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig. 7 Olaparib induces stromal fibroblast activation in human OC tumor specimens. a Immunohistochemical detection of alpha-SMA in matched tumor specimens from patients with OC before and after administration with Ola. Scale bar, 50 um. b Quantification of alpha-SMA-positive areas in tumor specimens from OC patients. Same images were measured repeatedly. c - e Representative images and quantification of Masson's trichrome and picrosirius red staining showing that Ola administration increased the content of stromal components and collagen deposition in human OC tumor specimens. Scale bar, 50 um. Same images were measured repeatedly. f , g Representative IHC images and quantification of CCL5 and p-P65 in matched tumor specimens from patients with OC before and after administration with Ola. Scale bar, 50 um. Measures were taken from different samples ( n = 6). h Correlation analysis of CCL5 IHC scores and alpha-SMA IHC scores in matched tumor specimens from patients with OC before and after administration with Ola. i Correlation analysis of p-P65 IHC scores and CCL5 IHC scores in matched tumor specimens from patients with OC before and after administration with Ola. j Schematic representation of the effect of PARPis on human ovarian stromal fibroblasts. Data are expressed as mean +- s.e.m., * p < 0.05; ** p < 0.01; *** p < 0.001.