AIF-1/Iba1 Antibody

Novus Biologicals | Catalog # NB100-2833

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse

Cited:

Mouse

Predicted:

Canine (100%). Backed by our 100% Guarantee.

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Peptide ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence

Cited:

Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, IF/IHC

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
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Product Specifications

Immunogen

This AIF-1/Iba1 Antibody was developed against a peptide with sequence C-NKQFLDDPKYSSDED corresponding to internal region according to NP_001614.3.

Reactivity Notes

Use in Mouse reported in scientific literature (PMID:31560162).

Marker

pan-Microglia Marker

Specificity

This AIF-1/Iba1 Antibody is expected to recognize isoform 3 (NP_001614.3) only.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Theoretical MW

16.7 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for AIF-1/Iba1 Antibody

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833]

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833]

Immunocytochemistry/Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] - Immunofluorescence analysis of paraformaldehyde fixed U937 cells immobilized on Shi-fix coverslip, permeabilized with 0.15% Triton. Primary incubation 1hr (10ug/ml) followed by Alexa Fluor 488 secondary antibody (2ug/ml), showing cytoplasmic staining. The nuclear stain is DAPI (blue). Negative control: Unimmunized goat IgG (10ug/ml) followed by Alexa Fluor 488 secondary antibody (2ug/ml).
Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody [NB100-2833]

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody [NB100-2833]

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody [NB100-2833] - Staining (6ug/ml) of paraffin embedded Human Lung. Heat induced antigen retrieval with citrate buffer pH 6, HRP-Staining.
Flow Cytometry: AIF-1/Iba1 Antibody [NB100-2833]

Flow Cytometry: AIF-1/Iba1 Antibody [NB100-2833]

Flow Cytometry: AIF-1/Iba1 Antibody [NB100-2833] - Flow cytometric analysis of paraformaldehyde fixed K562 cells (blue line), permeabilized with 0.5% Triton. Primary incubation 1hr (10ug/ml) followed by Alexa Fluor 488 secondary antibody (1ug/ml). IgG control: Unimmunized goat IgG (black line) followed by Alexa Fluor 488 secondary antibody.
Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833]

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833]

AIF-1-Iba1-Antibody-Immunocytochemistry-Immunofluorescence-NB100-2833-img0012.jpg
Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833]

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833]

Immunocytochemistry/Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] - Immunofluorescence analysis of paraformaldehyde fixed CaCo-2 cells immobilized on Shi-fix coverslip, permeabilized with 0.15% Triton. Primary incubation 1hr (10ug/ml) followed by Alexa Fluor 488 secondary antibody (2ug/ml), showing cytoplasmic staining. The nuclear stain is DAPI (blue). Negative control: Unimmunized goat IgG (10ug/ml) followed by Alexa Fluor 488 secondary antibody (2ug/ml).
Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody [NB100-2833]

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody [NB100-2833]

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody [NB100-2833] - Negative Control showing staining of paraffin embedded Human Lung, with no primary antibody.
AIF-1/Iba1 Antibody

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] -

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] - Pg/gingipain is present intracellularly in astrocytes, microglia & neurons in the hippocampus of experimental mice.Green: astrocytes (top panels), microglia (middle panels), & neurons (bottom panels). Red: Pg/gingipain. Blue: DAPI. Representative of N = 4 (GFAP), 3 (Iba1) & 4 (NeuN) experimental mice. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30281647), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
AIF-1/Iba1 Antibody

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] -

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] - Proinflammatory cytokines IL6, IL1 beta & TNF alpha are evident in the hippocampus of experimental but not in control mice.Data from IF microscopy & RT-PCR. (A-F) Results from IF microscopy. IL6 expression in control (A) & experimental (B) mice, IL1 beta expression in control (C) & experimental (D) mice & TNF alpha expression in control (E) & experimental (F) mice. N = 3 mice/group. (G-I) Gene expression of cytokines was detected by RT-PCR (G, H, I). In all cases, there is significantly higher gene expression of all three cytokines in experimental compared with control group (G, H, I). Green: Iba1, Red: cytokines, Blue: DAPI. N = 5 mice/group. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30281647), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
AIF-1/Iba1 Antibody

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] -

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] - Proinflammatory cytokines IL6, IL1 beta & TNF alpha are evident in the hippocampus of experimental but not in control mice.Data from IF microscopy & RT-PCR. (A-F) Results from IF microscopy. IL6 expression in control (A) & experimental (B) mice, IL1 beta expression in control (C) & experimental (D) mice & TNF alpha expression in control (E) & experimental (F) mice. N = 3 mice/group. (G-I) Gene expression of cytokines was detected by RT-PCR (G, H, I). In all cases, there is significantly higher gene expression of all three cytokines in experimental compared with control group (G, H, I). Green: Iba1, Red: cytokines, Blue: DAPI. N = 5 mice/group. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30281647), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
AIF-1/Iba1 Antibody

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] -

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] - IL-1 beta synthesis, & microglial activation modulation in PDD005-treated aged mice. (A) Images showing IL-1 beta (green fluorescence), iba-1 (red fluorescence) expression. Yellow = Overlay; Blue = nuclei; SGZ region of mouse brain. 40X magnification, scale bar = 100 μm. (B) Graphs representing the average relative fluorescence of Iba-1 in SGZ. Significant differences determined by using one-way ANOVA with Tukey’s test. *P < 0.050, **P < 0.01 & ***P < 0.001 compared with aged-vehicle. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31980673), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
AIF-1/Iba1 Antibody

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] -

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] - Proinflammatory cytokines IL6, IL1 beta & TNF alpha are evident in the hippocampus of experimental but not in control mice.Data from IF microscopy & RT-PCR. (A-F) Results from IF microscopy. IL6 expression in control (A) & experimental (B) mice, IL1 beta expression in control (C) & experimental (D) mice & TNF alpha expression in control (E) & experimental (F) mice. N = 3 mice/group. (G-I) Gene expression of cytokines was detected by RT-PCR (G, H, I). In all cases, there is significantly higher gene expression of all three cytokines in experimental compared with control group (G, H, I). Green: Iba1, Red: cytokines, Blue: DAPI. N = 5 mice/group. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30281647), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
AIF-1/Iba1 Antibody

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] -

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] - Proinflammatory cytokines IL6, IL1 beta & TNF alpha are evident in the hippocampus of experimental but not in control mice.Data from IF microscopy & RT-PCR. (A-F) Results from IF microscopy. IL6 expression in control (A) & experimental (B) mice, IL1 beta expression in control (C) & experimental (D) mice & TNF alpha expression in control (E) & experimental (F) mice. N = 3 mice/group. (G-I) Gene expression of cytokines was detected by RT-PCR (G, H, I). In all cases, there is significantly higher gene expression of all three cytokines in experimental compared with control group (G, H, I). Green: Iba1, Red: cytokines, Blue: DAPI. N = 5 mice/group. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30281647), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
AIF-1/Iba1 Antibody

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] -

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] - Proinflammatory cytokines IL6, IL1 beta & TNF alpha are evident in the hippocampus of experimental but not in control mice.Data from IF microscopy & RT-PCR. (A-F) Results from IF microscopy. IL6 expression in control (A) & experimental (B) mice, IL1 beta expression in control (C) & experimental (D) mice & TNF alpha expression in control (E) & experimental (F) mice. N = 3 mice/group. (G-I) Gene expression of cytokines was detected by RT-PCR (G, H, I). In all cases, there is significantly higher gene expression of all three cytokines in experimental compared with control group (G, H, I). Green: Iba1, Red: cytokines, Blue: DAPI. N = 5 mice/group. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30281647), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
AIF-1/Iba1 Antibody

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] -

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] - Proinflammatory cytokines IL6, IL1 beta & TNF alpha are evident in the hippocampus of experimental but not in control mice.Data from IF microscopy & RT-PCR. (A-F) Results from IF microscopy. IL6 expression in control (A) & experimental (B) mice, IL1 beta expression in control (C) & experimental (D) mice & TNF alpha expression in control (E) & experimental (F) mice. N = 3 mice/group. (G-I) Gene expression of cytokines was detected by RT-PCR (G, H, I). In all cases, there is significantly higher gene expression of all three cytokines in experimental compared with control group (G, H, I). Green: Iba1, Red: cytokines, Blue: DAPI. N = 5 mice/group. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30281647), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
AIF-1/Iba1 Antibody

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] -

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] - Lifelong choline supplementation alters the expression of the alpha7 nicotinic acetylcholine receptor ( alpha 7nAchR) within microglia. (a) Photomicrographs depicting the Cornus Ammonis 1 (CA1) of the hippocampus from APP/PS1 & NonTg mice fluorescently stained for alpha 7nAchR & Iba1. Images taken at 40X; scale bar = 25 µm (n = 6 mice/group). (b) Quantitative analysis reveals a significant main effect of genotype, where the APP/PS1 mice have a significantly higher intensity of yellow pixels of alpha 7nAchR/Iba1 colocalization than the NonTg mice (p < .01). Additionally, we find a significant main effect of diet, where the Ch+ groups show a significant reduction in alpha 7nAchR/Iba1 colocalization than the CTL groups (p < .05). A genotype by diet interaction was found, where the APP/PS1 Ch+ mice show a significant reduction of alpha 7nR/Iba1 colocalization compared to the CTL counterparts (p < .001). The center line represents the median value, the limits represent the 25th & 75th percentile, & the whiskers represent the minimum & maximum value of the distribution. **p < .01, ***p < .001 Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31560162), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
AIF-1/Iba1 Antibody

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] -

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] - (A–D) Images of coronal brain sections showing DiI (red) labeled EVs taken up by IBA1 (blue) & CD11b (green) positive microglia. (E,F) Tomato & control plasmid (blue) dorsal SVZ electroporations stained for CD11b (green) & IBA1 (magenta) at P7, 10x (E), 20x (F). (G,H) 20x magnification of E showing CD11b (green) & IBA1 (red) (G) or CD11b (H). (I,J) Tomato & Rab27a shRNA plasmid (blue) dorsal SVZ electroporations stained for CD11b (green) & IBA1 (magenta) at P7, 10x (I), 20x (J). (K,L) 20x magnification of I showing CD11b (green) & IBA1 (red) (K) or CD11b (L). (M) Western blot of Rab27a following shRNA mediated knockdown. (N) Images of coronal section stained for Rab27a (green) following control or Rab27a shRNA & tomato (magenta) co-electroporation at P0 & sacrificed 48 hrs later. (O) Quantification of CD11b positive microglia at P2 & P7 following control or Rab27a shRNA electroporation. Data are represented as mean ± SEM. ****p < 0.0001 A, E, I scale bar, 100 μm. B–D, F–H, J–L, N scale bar, 50 μm. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30816224), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
AIF-1/Iba1 Antibody

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] -

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] - Lifelong choline supplementation reduces activated microglia. (a) Photomicrographs depicting the Cornus Ammonis 1 (CA1) of the hippocampus from APP/PS1 & NonTg mice fluorescently stained for CD68 & Iba1. Images taken at 10X; Scale bar = 150 µm (n = 6 mice/group). Arrows illustrate colocalization of CD68/Iba1. (b) Quantitative analysis reveals a significant main effect of genotype, where the APP/PS1 mice have a significantly higher intensity of yellow pixels of CD68/Iba1 colocalization than the NonTg mice (p < .05). Additionally, we find a significant main effect of diet, where the Ch+ groups show a significant reduction in CD68/Iba1 colocalization than the CTL groups (p < .001). Data are presented as box plots. The center line represents the median value, the limits represent the 25th & 75th percentile, & the whiskers represent the minimum & maximum value of the distribution. *p < .05, ***p < .001 Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31560162), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
AIF-1/Iba1 Antibody

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] -

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody [NB100-2833] - Lifelong choline supplementation alters the expression of the Sigma‐1 receptor ( sigma 1R) within microglia. (a‐b) Representative Western blot of sigma 1R levels. Quantitative analysis of sigma 1R protein levels reveals a significant reduction with Ch+ (p < .05; n = 5/APP group; n = 4/NonTg group). (c‐d) Quantitative analysis reveals a significant main effect of genotype, where the APP/PS1 mice have a significantly higher intensity of yellow pixels of sigma 1R/Iba1 colocalization than the NonTg mice (p < .05, n = 6 APP/PS1 CTL, n = 5 APP/PS1 Ch+, n = 6 NonTg CTL, n = 6 NonTg Ch+). Additionally, we find a significant genotype by diet interaction where the APP/PS1 Ch+ mice show a significant reduction in sigma 1R/Iba1 colocalization than the APP/PS1 CTL mice (p < .001). Photomicrographs depicting the Cornus Ammonis 1 (CA1) of the hippocampus from APP/PS1 & NonTg mice fluorescently stained for the  sigma 1R & Iba1; images taken at 40×; scale bar = 25 µm. Data are presented as box plots. The center line represents the median value, the limits represent the 25th & 75th percentile, & the whiskers represent the minimum & maximum value of the distribution. *p < .05, ***p < .001 Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31560162), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
AIF-1/Iba1 Antibody

Western Blot: AIF-1/Iba1 Antibody [NB100-2833] -

Western Blot: AIF-1/Iba1 Antibody [NB100-2833] - Astrocytic marker was decreased in the brain of prenatally saline-exposed offspring, but not in the offspring prenatally exposed to LPS, following pharmacological activation of mGluR5.A prenatal administration of LPS did not change the brain expression level of iba1, CD68, GFAP or mGluR5 in 4-month-old mice (A). Postnatal CDPPB treatment reduced the GFAP level, without effect on iba1, CD68 or mGluR5 in the mice prenatally exposed to saline solution. The quantified molecular markers did not change in the brain of mice treated with MTEP (B). No change in iba1, CD68, GFAP or mGluR5 was observed in the brain of the mice prenatally exposed to LPS (C). Values are expressed as mean ± SEM. Abbreviations: MTEP, 3-((2-methyl-4-thiazolyl)ethynyl)pyridine; CDPPB, 3-cyano-N-(1,3-diphenyl-1H-pyrazol-5-yl) benzamide; CD68, cluster of differentiation 68); iba1, ionized calcium binding adaptor molecule-1; GFAP, glial fibrillary acidic protein; mGluR5, metabotropic glutamate receptor subtype 5; ROD, relative optical density. **p < 0.01 >> Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26536027), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for AIF-1/Iba1 Antibody

Application
Recommended Usage

Flow Cytometry

10 ug/mL

Immunocytochemistry/ Immunofluorescence

10 ug/mL

Immunohistochemistry-Paraffin

6 ug/ml

Peptide ELISA

Detection limit 1:128000
Application Notes
Use in Immunohistochemistry reported in scientific literature (PMID:31560162).

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Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

Tris saline (20 mM Tris pH 7.3, 150 mM NaCl), 0.5% BSA

Preservative

0.02% Sodium Azide

Concentration

0.5 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C. Avoid freeze-thaw cycles.

Background: AIF-1/Iba1

Allograft inflammatory factor (AIF-1) or ionized calcium-binding adapter molecule 1 (Iba1) is a cytosolic actin binding protein containing a calcium binding domain (EF-hand) and inducible by cytokines and IFN-gamma (1). AIF-1 (theoretical molecular weight 17kDa) was first cloned from rat and human cardiac allografts and macrophage cell lines. In cardiac allografts, expression of AIF-1 was specifically associated with infiltrated mononuclear cells (2, 3). AIF-1 is constitutively expressed and inducible in macrophages and microglia. Iba1, microglia response factor (MRF-1) and daintain were independently cloned and identified in rat and human tissues and share complete sequence identity with AIF-1. Additionally, several AIF-1 splice variants have been identified including IRT-1, BART-1, G1 and Hara-1 (2).

Several cellular functions have been associated with AIF-1/Iba1 expression including cell growth, cell migration, actin bundling, membrane ruffling, and phagocytic activity (2, 4). Iba1 induces Rac signaling through a PLC-gamma dependent pathway (1). Rac, a member of the Rho family of small GTPases, localizes with Iba1 and F-actin in membrane ruffles and phagocytic cups and plays a role in microglia activation. AIF-1/Iba1 induction in macrophages and microglia occur in association with immunological inflammatory processes in various disease states including endometriosis, cerebral infarction and rheumatoid arthritis (5). Immunodetection of Iba1 through flow cytometry, immunohistochemical or immunocytochemical applications is commonly used for identification and analysis of microglia.

References

1. Imai, Y., & Kohsaka, S. (2002). Intracellular signaling in M-CSF-induced microglia activation: Role of Iba1. GLIA. https://doi.org/10.1002/glia.10149

2. Deininger, M. H., Meyermann, R., & Schluesener, H. J. (2002). The allograft inflammatory factor-1 family of proteins. FEBS Letters. https://doi.org/10.1016/S0014-5793(02)02430-4

3. Utans, U., Quist, W. C., Mcmanus, B. M., Wilson, J. E., Arceci, R. J., Wallace, A. F., & Russell, M. E. (1996). Allograft inflammatory factor-1: A cytokine-responsive macrophage molecule expressed in transplanted human hearts. Transplantation. https://doi.org/10.1097/00007890-199605150-00018

4. Franco-Bocanegra, McAuley, Nicoll, & Boche. (2019). Molecular Mechanisms of Microglial Motility: Changes in Ageing and Alzheimer's Disease. Cells. https://doi.org/10.3390/cells8060639

5. Kimura, M., Kawahito, Y., Obayashi, H., Ohta, M., Hara, H., Adachi, T.,... Yoshikawa, T. (2007). A Critical Role for Allograft Inflammatory Factor-1 in the Pathogenesis of Rheumatoid Arthritis. The Journal of Immunology. https://doi.org/10.4049/jimmunol.178.5.3316

Long Name

Allograft Inflammatory Factor 1

Alternate Names

AIF1, IBA1, IRT1

Entrez Gene IDs

199 (Human); 11629 (Mouse)

Gene Symbol

AIF1

Additional AIF-1/Iba1 Products

Product Documents for AIF-1/Iba1 Antibody

Certificate of Analysis

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Product Specific Notices for AIF-1/Iba1 Antibody

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for AIF-1/Iba1 Antibody

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